KR20210116548A - PCSK9 inhibitors and methods of use thereof - Google Patents

Abstract

The present invention relates to novel heteroaryl compounds and pharmaceutical formulations thereof. The present invention also relates to a method for the treatment or prevention of cardiovascular disease, and a method for the treatment of sepsis or septic shock, using the novel heterocyclic compounds described herein.

Description

PCSK9 inhibitors and methods of use thereof

[Related Application]

This application claims the benefit of priority from US Application No. 62/794,234, filed on January 18, 2019, which is incorporated herein by reference in its entirety.

PCSK9, also referred to as "preprotein convertase subtilisin/kexin 9", is a member of the secretory proprotein convertase family and plays an important role in cholesterol metabolism. PCSK9 increases the level of circulating LDL cholesterol through enhanced degradation of LDL receptors independent of catalytic activity. Secreted PCSK9 binds to the epidermal growth factor domain A (EGFA) of the LDL receptor (LDLR) at the cell surface and the PCSK9/LDL receptor complex is internalized into the endosomal/lysosomal compartment. The enhanced binding affinity of PCSK9 to the LDL receptor at the acidic pH of the late endosome/lysosome reduces recycling of the LDL receptor and instead targets the LDL receptor for lysosomal degradation. Genetic linkage studies have demonstrated that loss-of-function mutations in PCSK9 are associated with lower plasma LDL-C levels and reduced incidence of adverse cardiovascular events.

Another biological pathway implicated in the effect of PCSK9 on the LDL receptor is the pathogenesis of septic shock. Septic shock is an often fatal complication of a severe microbial infection (sepsis) that causes an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis occurs in microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharides (LPS; Gram-negative bacteria). Since LPS is a potent ligand for mammalian innate immune receptors (Toll-like receptors (TLRs)), it is prominent in the septic inflammatory response (septic shock or sepsis).

PCSK9 reduces LPS uptake by the liver's LDL receptors, causing free LPS to overstimulate the body's immune response to the pathogens that cause sepsis. Thus, inhibiting PCSK9 is beneficial in maintaining hepatic LDL receptors to achieve systemic pathogen clearance and detoxification for sepsis. However, there is currently no effective treatment for sepsis or septic shock other than antibiotic therapy.

For cardiovascular disease, there are few options for inhibiting PCSK9. Statins actually up-regulate PCSK9 in HepG2 cells and human primary hepatocytes through increased expression of SREBP-2, a transcription factor that up-regulates both LDLR and PCSK9 genes. Since increased levels of PCSK9 reduce the amount of LDL receptors on the cell surface, a proportional LDL-cholesterol lowering effect could not be achieved by increasing the statin dose.

Alirocumab and evolocumab, two monoclonal antibodies (mAbs) that selectively bind extracellular PCSK9 and prevent interaction with the LDL receptor, have recently received FDA approval for lowering LDL-C levels. In clinical trials, alirocumab showed an approximately 50% reduction in LDL levels compared to placebo. Elbitar et al., Expert Opin Therapeutic Patents 2016 26:1377-1392. Patients taking evolocumab showed a reduction of about 60-75% in LDL levels. The efficacy of these drugs shows the potential for PCSK9 inhibitors to be an effective treatment for patients with hypercholesterolemia and other cardiovascular diseases. However, both antibody drugs require intravenous administration and can cause allergic or other adverse immune reactions in the body.

Cardiovascular disease often requires lifelong management, unlike infections that can be transient. Thus, ease of dosing and administration is a key factor for patient compliance with maintenance medication. There is a need for PCSK9 inhibitors with increased potency and greater ease of administration that can be achieved with small molecule PCSK9 inhibitors.

Disclosed herein are compounds of formula (I),

[Formula I]

In the chemical formula,

A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR ⁶ , and —C(O)NR ⁶ R ⁷ ;

B is selected from H, alkyl, and halo, or

A and B together with the carbon atom to which they are attached form a 5 or 6 membered heteroaryl;

X is NR ⁵ or O;

R ¹ and R ^1′ are each independently selected from H and alkyl; or

when n is 0, R ¹ and R ^1′ together with the atoms to which they are attached form a 4 to 8 membered cycloalkyl or cycloalkenyl ring;

R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring; or

R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ^2′ is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano, or

R ² and R ^2′ together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

each of R ³ and R ⁴ is independently H or alkyl; or

R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ⁵ is H or alkyl; or

R ¹ and R ⁵ together with the atoms to which they are attached form a 6 to 8 membered cycloalkyl or heterocyclyl ring; or

R ² and R ⁵ together with the atoms to which they are attached form a 5 to 8 membered cycloalkyl or heterocyclyl ring;

each R ⁶ and R ⁷ is independently H or alkyl;

Y is selected from aryl, heteroaryl and heterocyclyl;

n is 0 or 1.

In certain embodiments, the present invention provides treatment for cardiovascular disease comprising an effective amount of any compound described herein (eg, a compound of the present invention, such as a compound of Formula I), and one or more pharmaceutically acceptable excipients. or a pharmaceutical composition suitable for use in a subject in prophylaxis. In certain embodiments, the pharmaceutical formulation may be for use in treating or preventing a condition or disease as described herein.

Disclosed herein are methods of treating diseases and conditions that would benefit from inhibition of PCSK9. These diseases include cardiovascular diseases such as hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome and coronary artery disease. It is not limited thereto.

Other diseases and conditions that can be treated using the methods described herein include, but are not limited to, sepsis and septic shock.

Provided herein are combination therapies of a compound of formula (I) with monoclonal antibodies, statins and other cardiovascular agents that may enhance cardiovascular therapeutic benefits beyond the ability of adjuvant therapy alone. Also provided herein is a combination therapy of a compound of formula (I) with an antibiotic capable of reducing the incidence and severity of sepsis and septic shock beyond the ability of adjuvant therapy alone.

Disclosed herein are compounds of formula (I),

[Formula I]

In the chemical formula,

A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR ⁶ , and —C(O)NR ⁶ R ⁷ ;

B is selected from H, alkyl, and halo, or

A and B together with the carbon atom to which they are attached form a 5 or 6 membered heteroaryl;

X is NR ⁵ or O;

R ¹ and R ^1′ are each independently selected from H and alkyl; or

when n is 0, R ¹ and R ^1′ together with the atoms to which they are attached form a 4 to 8 membered cycloalkyl or cycloalkenyl ring;

R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring; or

R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ^2′ is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano, or

R ² and R ^2′ together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

each of R ³ and R ⁴ is independently H or alkyl; or

R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ⁵ is H or alkyl; or

R ¹ and R ⁵ together with the atoms to which they are attached form a 6 to 8 membered cycloalkyl or heterocyclyl ring; or

R ² and R ⁵ together with the atoms to which they are attached form a 5 to 8 membered cycloalkyl or heterocyclyl ring;

each R ⁶ and R ⁷ is independently H or alkyl;

Y is selected from aryl, heteroaryl and heterocyclyl;

n is 0 or 1.

Disclosed herein are compounds of Formula I',

[Formula I']

In the chemical formula,

A is selected from H, halo, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR ⁶ , and —C(O)NR ⁶ R ⁷ ;

B is selected from H, alkyl, and halo;

R ¹ and R ^1′ are each independently selected from H and alkyl; or

when n is 0, R ¹ and R ^1′ together with the atoms to which they are attached form a 4 to 8 membered cycloalkyl ring;

R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring; or

R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ^2′ is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano; or

R ² and R ^2′ together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

each of R ³ and R ⁴ is independently H or alkyl; or

R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;

R ⁵ is H or alkyl; or

R ¹ and R ⁵ together with the atoms to which they are attached form a 6 to 8 membered cycloalkyl or heterocyclyl ring; or

R ² and R ⁵ together with the atoms to which they are attached form a 5 to 8 membered cycloalkyl or heterocyclyl ring;

each R ⁶ and R ⁷ is independently H or alkyl;

Het is heteroaryl or heterocyclyl;

n is 0 or 1.

All of the embodiments below and herein are to be understood as embodiments of both Formula I and Formula I′.

In certain embodiments, according to claim 1, A is H, hydroxy, thioalkyl, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR ⁶ , and -C(O)NR ⁶ R ⁷ .

In certain embodiments, A is H, but in other embodiments, A is alkyl, such as thioalkyl. In certain embodiments, A is selected from -SCH ₃ , -SCHF ₂ , and -OCHF ₂ . In some embodiments, A is alkoxy. In other embodiments, A is cycloalkyl. In certain embodiments, B is H.

In certain embodiments, A and B together with the carbon atom to which they are attached form a pyrrolyl or thienyl ring, which is unsubstituted or substituted with one or more alkyl.

In certain embodiments, X is preferably NR5. In other embodiments, Y is preferably heteroaryl or heterocyclyl.

In certain embodiments, R ¹ and R ¹ ′ are each H. However, when n is 0, R ¹ and R ^1′ together with the atoms to which they are attached may form a 4 to 8 membered cycloalkyl ring. In some embodiments, cycloalkyl is monocyclic or bicyclic. In other embodiments, R ¹ and R ^1′ together with the atoms to which they are attached may form a 4 to 8 membered cycloalkenyl ring. In some embodiments, the cycloalkyl ring is a cyclopentyl ring, such as S,S -cyclopentyl. In some embodiments, the cycloalkyl ring is substituted with hydroxyl or hydroxyalkyl.

In certain embodiments, R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, alkylamino, cyano, and hydroxyl. In certain embodiments, R ² is C _1-3 alkyl. R ² may be substituted with one or more substituents selected from amino, amido, cyano, hydroxy, and heterocyclyl. In some embodiments, R ^2′ is C _1-3 alkyl, while in other embodiments, R ^2′ is H.

In certain embodiments, R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. In other embodiments, R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. In certain embodiments, R ² and R ^2′ together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring.

In certain embodiments, R ³ is C _1-3 alkyl, while in other embodiments, R ³ is H. In certain embodiments, R ⁴ is H.

In certain embodiments, R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. In other embodiments, R ¹ and R ⁵ together with the atoms to which they are attached form a 6-8 membered cycloalkyl or heterocyclyl ring. In other embodiments, R ² and R ⁵ together with the atoms to which they are attached form a 5-8 membered cycloalkyl or heterocyclyl ring.

In certain embodiments, Y is monocyclic heteroaryl, such as, but not limited to, pyridinyl, pyrazinyl, pyrimidinyl, and thiazolyl. In other embodiments, Y is monocyclic heteroaryl, such as, but not limited to, pyridinyl, pyrazinyl, and pyrimidinyl. In some embodiments, Y is selected from trizenyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl and triazolyl. monocyclic heteroaryl may be unsubstituted or 1 selected from alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, heteroaryl, nitro, sulfonamido, and thioalkyl It may be substituted with more than one substituent. In another embodiment, Y is monocyclic heteroaryl, which may be unsubstituted, or alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, aryl, heteroaryl, heterocycle and may be substituted with one or more substituents selected from ryl, nitro, sulfonamido, and thioalkyl.

In certain embodiments, monocyclic heteroaryl is phenyl, pyridinyl, 2-hydroxypyridinyl, piperidinonyl, 2-hydroxy-1-methylpyridinyl, triazolyl, imidazolidinonyl, pyrimidonyl , aryl, heteroaryl or heterocyclyl selected from 2-hydroxyisoquinolinyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinonyl. In certain preferred embodiments, Y is 6 membered monocyclic heteroaryl. In some embodiments, monocyclic heteroaryl is substituted with heteroaryl or heterocyclyl substituted with one or more substituents selected from halo, CN, alkyl, alkoxy, hydroxy, carboxy, —CO _{2 alkyl, and tetrazolyl.} In certain preferred embodiments, monocyclic heteroaryl is placed in the para position of A with respect to X.

In other embodiments, Y is bicyclic heteroaryl such as, but not limited to, benzothiazolyl, benzimidazolyl, benzoxazolyl, triazolopyridinyl, thiazolopyridinyl, quinolinyl, and quinoxali it's neil Bicyclic heteroaryl may be unsubstituted or selected from alkyl, haloalkyl, hydroxyalkyl, thioalkyl, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamido may be substituted with one or more substituents. In certain embodiments, bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from thioalkyl, alkoxycarbonyl, amido, carboxy, halo, and heteroaryl.

In certain embodiments, Y is substituted with an amido substituent of the formula -C(O)NR ⁸ R ⁹ or -NR ⁹ C(O)R ^{10 , wherein:}

R ⁸ and R ⁹ are each independently selected from H, alkyl, heterocyclyl and heteroaryl; or

R ⁸ and R ⁹ together with the nitrogen atom to which they are attached form a 4, 5, 6, or 7 membered heterocyclic or heteroaryl ring;

R ¹⁰ is alkyl.

In certain embodiments, Y is substituted with a sulfonamido substituent of the formula -S(O) ₂ NR ⁸ R ⁹ or -NR ⁹ S(O) ₂ R ^{10 , wherein}

R ⁸ and R ⁹ are each independently selected from H, alkyl, and heteroaryl; or

R ⁸ and R ⁹ together with the nitrogen atom to which they are attached form a 4, 5, 6, or 7 membered heterocyclic ring;

R ¹⁰ is alkyl.

It should be understood that all the aforementioned embodiments of the variable Y of formula I are also embodiments of the variable Het of formula I'.

In certain embodiments, R ⁸ and R ⁹ are each independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl. In ^{embodiments wherein one or both of R 8} and R ⁹ are alkyl, each alkyl is independently unsubstituted, or methyl, methoxy, carboxy, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, Methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-dihydro-3H-1,2,4-tria substituted with one or more substituents selected from zol-3-onyl. In certain embodiments, R ⁸ and R ⁹ together with the nitrogen atom to which they are attached are aziradine, isothiazolidin-1,1-dioxide, azetidine, thiazol-4(5Hn)-one, morpholine, p form a heterocyclic ring selected from peridine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane. In some embodiments, R ⁸ and R ⁹ together with the nitrogen atom to which they are attached are 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine, octahydropyrazino [2,1-c] [1,4] oxazine, tetrahydropyrido [3,4-d] pyrimidine, 2-oxa-8-azaspiro [4.5] decane, tetrahydropyrrolo [3,4 -c] pyrazole, thiomorpholine, 2-oxa-7-azaspiro [3.5] nonane, 2,8-diazaspiro [4.5] decan-3-one, tetrahydro-1,7-naphthyridine, 1 -oxa-4,9-diazaspiro[5.5]undecan-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, Hexahydro-3H-oxazolo[3,4-a]pyrazin-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro-[1,2, 4] triazolo [4,3-a] pyrazine, 2,7-diazaspiro [4.4] nonane, 2,6-diazaspiro [3.4] octane, 7-oxa-2-azaspiro [3.5] nonane, 1-oxa-8λ ² -azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridinone, tetrahydro-[1,2,4 ]triazolo[4,3-a]pyrazin-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5 ] form a heterocyclic ring selected from nonane, pyrazole, and pyridazin-3(2H)-one.

In some embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl. In certain embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxy, and oxetanyl.

In certain embodiments, the invention provides a pharmaceutical formulation suitable for use in a human subject comprising any of the compounds set forth above (e.g., a compound of the invention, such as a compound of Formula I, and one or more pharmaceutically acceptable excipients). In certain embodiments, the pharmaceutical formulation may be for use in treating or preventing a condition or disease as described herein.

Any of the compounds disclosed may be used in the manufacture of a medicament for the treatment of any disease or condition disclosed herein.

Justice

Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art of this disclosure. The following references provide those skilled in the art with general definitions for many of the terms used in this disclosure: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991)]; and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings assigned to them below, unless otherwise specified.

In this disclosure, "comprises", "comprising", "comprising may mean "include", "including" and the like; "Consisting essentially of" or "consisting essentially of" likewise has the meaning given to U.S. patent law, which term means unless the basic or novel features of the enumerated are altered by existence beyond the enumerated; It is open-ended which allows for more than enumerated, but excludes prior art implementations.

Unless specifically stated or clear from the context, the term “or” as used herein is to be understood as inclusive. As used herein, the terms “a”, “an”, and “the” are to be understood in the singular or plural, unless specifically stated or clear from the context.

The term "acyl" is known in the art and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

The term “acylamino” is known in the art and refers to an amino group substituted with an acyl group, and may be represented, for example, by the formula hydrocarbylC(O)NH—.

The term "acyloxy" is known in the art and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

The term “alkoxy” refers to an alkyl group to which an oxygen is attached, preferably a lower alkyl group. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

As used herein, the term “alkenyl” refers to an aliphatic group containing at least one double bond and is intended to include both “unsubstituted alkenyl” and “substituted alkenyl,” the latter of which is one of the alkenyl groups. Refers to an alkenyl moiety having a substituent replacing a hydrogen on one or more carbons. Such substituents may occur at one or more carbons included or not included in one or more double bonds. Moreover, such substituents include all contemplated for alkyl groups as discussed below, except where stability is prohibited. For example, substitution of an alkenyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

An “alkyl” group or “alkane” is a fully saturated straight-chain or branched non-aromatic hydrocarbon. Typically, straight-chain or branched alkyl groups have from 1 to about 20 carbon atoms, preferably from 1 to about 10 carbon atoms, unless otherwise defined. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl and octyl. C ₁ -C ₆ straight or branched chain alkyl groups are also referred to as “lower alkyl” groups.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout the specification, examples and claims is intended to include both “unsubstituted alkyl” and “substituted alkyl,” the latter of which is one of the hydrocarbon backbone. refers to an alkyl moiety having a substituent replacing a hydrogen on one or more carbons. Such substituents, unless otherwise specified, are, for example, halogen, hydroxyl, carbonyl (eg, carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (eg, thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that a moiety substituted on the hydrocarbon chain may itself be substituted where appropriate. For example, the substituents of a substituted alkyl include amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamidos, sulfamoyl and sulfonates). , and substituted and unsubstituted forms of silyl groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxylates and esters), -CF ₃ , -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyl may be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl substituted alkyl, -CF ³ , -CN, and the like.

_{The term “C xy} ” when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups containing x to y carbons in the chain. For example, the term "C _xy alkyl" refers to straight and branched chain alkyl groups containing x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, and the like. substituted or unsubstituted saturated hydrocarbon groups, including C ₀ alkyl represents hydrogen when the group is in a terminal position and a bond when internal to the group. The terms “C _2-y alkenyl” and “C _2-y alkynyl” refer to substituted or unsubstituted unsaturated aliphatic groups of similar length and capable of substitution with the aforementioned alkyls, but each containing at least one double or triple bond. refers to

As used herein, the term “alkylamino” refers to an amino group substituted with at least one alkyl group.

As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

As used herein, the term “alkynyl” refers to an aliphatic group containing at least one triple bond and is intended to include both “unsubstituted alkynyl” and “substituted alkynyl,” the latter of which is one of the alkynyl groups. Refers to an alkynyl moiety having a substituent replacing a hydrogen on one or more carbons. Such substituents may occur at one or more carbons included or not included in one or more triple bonds. Moreover, such substituents include all contemplated for alkyl groups as discussed above, except where stability is prohibited. For example, substitution of an alkynyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

As used herein, the term "amide" refers to the group

wherein each R ¹¹ independently represents a hydrogen or hydrocarbyl group, or two R ¹¹ together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

The terms "amine" and "amino" are known in the art and refer to unsubstituted and substituted amines and salts thereof, for example, moieties that may be represented by the formula

또는

or

wherein each R ¹¹ independently represents a hydrogen or hydrocarbyl group, or two R ¹¹ together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure. As used herein, the term “aminoalkyl” refers to an alkyl group substituted with an amino group.

As used herein, the term “aralkyl” refers to an alkyl group substituted with an aryl group.

As used herein, the term “aryl” includes substituted or unsubstituted single ring aromatic groups wherein each atom of the ring is carbon. Preferably, the ring is a 5 to 7 membered ring, more preferably a 6 membered ring. The term "aryl" also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjacent rings, wherein at least one of the rings is aromatic, e.g., the other ring is cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and/or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

The term "carbamate" is known in the art and refers to the group

In this case, R ¹¹ and R ¹² independently represent hydrogen or a hydrocarbyl group, for example, an alkyl group, or R ¹¹ and R ¹² together with the intervening atom(s) are a heterocycle having 4 to 8 atoms in the ring structure. complete the

As used herein, the terms “carbocyclic” and “carbocyclic” refer to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocyclic includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkane rings in which all carbon atoms are saturated and cycloalkene rings containing at least one double bond.

The term “carbocycle” includes 5 to 7 membered monocyclic and 8 to 12 membered bicyclic rings. Each ring of the bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring of the fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In exemplary embodiments, an aromatic ring, such as phenyl, may be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct -3-ene, naphthalene and adamantane. Exemplary fused carbocycles are decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene and bicyclo[4.1 .0]hept-3-ene. A “carbocycle” may be substituted at any one or more positions that may contain a hydrogen atom.

A “cycloalkyl” group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, monocyclic cycloalkyl groups have from 3 to about 10 carbon atoms, more typically from 3 to 8 carbon atoms, unless otherwise defined. The second ring of the bicyclic cycloalkyl may be selected from saturated, unsaturated and aromatic rings. Cycloalkyl includes bicyclic molecules in which one, two, or three or more atoms are shared between two rings. The term “fused cycloalkyl” refers to a bicyclic cycloalkyl in which each ring shares two adjacent atoms with the other ring. The second ring of the fused bicyclic cycloalkyl may be selected from saturated, unsaturated and aromatic rings. A “cycloalkenyl” group is a cyclic hydrocarbon containing one or more double bonds.

As used herein, the term “carbocyclylalkyl” refers to an alkyl group substituted with a carbocyclic group.

The term “carbonate” is known in the art and refers to the group _{—OCO 2} —R ^{10 , wherein R 10} represents a hydrocarbyl group.

As used herein, the term “carboxy” refers to a group represented by the _{formula —CO 2 H.}

As used herein, the term “ester” refers to the group ^{—C(O)OR 10 , wherein R 10} represents a hydrocarbyl group.

As used herein, the term “ether” refers to a hydrocarbyl group linked to another hydrocarbyl group through an oxygen. Thus, the ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be symmetric or asymmetric. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which may be represented by the general formula alkyl-O-alkyl.

As used herein, the terms “halo” and “halogen” mean halogen and include chloro, fluoro, bromo and iodo.

As used herein, the terms “hetaralkyl” and “heteroaralkyl” refer to an alkyl group substituted with a hetaryl group.

As used herein, the term “heteroalkyl” refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein the two heteroatoms are not adjacent.

The terms "heteroaryl" and "hetaryl" include a substituted or unsubstituted aromatic monocyclic structure, preferably a 5 to 7 membered ring, more preferably a 5 to 6 membered ring, the ring structure of which is at least 1 heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein at least one of the rings is heteroaromatic and For example, the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like.

As used herein, the term “heteroatom” refers to an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

The terms "heterocyclyl", "heterocycle" and "heterocyclic" refer to a substituted or unsubstituted non-aromatic ring structure, preferably a 3 to 10 membered ring, more preferably a 3 to 7 membered ring and its ring structure contains at least 1 heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein at least one of the rings is heterocyclic. and, for example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, and the like.

As used herein, the term “heterocyclylalkyl” refers to an alkyl group substituted with a heterocycle group.

The term "hydrocarbyl," as used herein, is bonded through a carbon atom that does not have an =O or =S substituent and typically has at least one carbon-hydrogen bond and a predominantly carbon backbone, but may optionally contain heteroatoms. refers to a device that has Thus, groups such as methyl, ethoxyethyl, 2-pyridyl and trifluoromethyl are considered hydrocarbyl for purposes herein, but acetyl (having an =O substituent at the linking carbon) and ethoxy (rather than carbon) connected via oxygen). Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

As used herein, the term “hydroxyalkyl” refers to an alkyl group substituted with a hydroxy group.

The term "lower" when used in conjunction with a chemical moiety, such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is intended to include groups having no more than 10, preferably no more than 6, non-hydrogen atoms in the substituent. will be. "Lower alkyl" refers to an alkyl group containing, for example, up to 10, preferably up to 6 carbon atoms. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents as defined herein are, for example, hydroxyalkyl and aralkyl (wherein, for example, an atom in an aryl group is a carbon in an alkyl substituent) (not counted when counting atoms), whether they appear alone or in combination with other substituents, are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl or lower alkoxy, respectively.

The terms “polycyclyl”, “polycyclic” and “polycyclic” refer to two or more rings in which two or more atoms are common to two adjacent rings, eg, the rings are “fused rings” (eg, cyclo alkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclyl). Each ring of the polycyclic may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic contains 3 to 10, preferably 5 to 7 atoms in the ring.

The term “silyl” refers to a silicon moiety to which three hydrocarbyl moieties are attached.

The term “substituted” refers to a moiety having a substituent replacing a hydrogen at one or more carbons of the backbone. "Substitution" or "substituted with" refers to a stable compound in which such substitution is dependent on the allowed valences of the atom and substituent substituted, and the substitution does not undergo transformation, e.g., spontaneously, e.g., by rearrangement, cyclization, removal, etc. It will be understood to include the implied clue of bringing. As used herein, the term “substituted” is intended to include all permissible substituents of organic compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents are one or more and may be the same or different for suitable organic compounds. For purposes of this invention, a heteroatom such as nitrogen may have a hydrogen substituent and/or any permissible substituent of the organic compounds described herein satisfying the valence of the heteroatom. A substituent may be any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (eg, carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (eg, thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that a substituent may itself be substituted where appropriate. Unless specifically stated as “unsubstituted,” reference to a chemical moiety herein is understood to include substituted variants. For example, reference to an “aryl” group or moiety includes implicitly both substituted and unsubstituted variants.

The term “sulfate” is known in the art _{and refers to the group —OSO 3} H or a pharmaceutically acceptable salt thereof.

The term "sulfonamide" is known in the art and refers to a group represented by the general formula:

In this case, R ¹¹ and R ¹² independently represent hydrogen or hydrocarbyl, for example alkyl, or R ¹¹ and R ¹² together with the intervening atom(s) represent a heterocycle having 4 to 8 atoms in the ring structure. complete

The term “sulfoxide” is known in the art and refers to the group ^{—S(O)—R 10 , wherein R 10} represents a hydrocarbyl group.

The term “sulfonate” is known in the art _{and refers to a SO 3} H group or a pharmaceutically acceptable salt thereof.

The term “sulfone” is known in the art and refers to the group _{—S(O) 2} —R ^{10 , wherein R 10} represents hydrocarbyl.

As used herein, the term “thioalkyl” refers to an alkyl group substituted with a thiol group.

As used herein, the term “thioester” refers to the group ^{—C(O)SR 10} or —SC(O)R ^{10 , wherein R 10} represents hydrocarbyl.

As used herein, the term “thioether” corresponds to an ether in which the oxygen has been replaced by sulfur.

The term "urea" is known in the art and can be represented by the general formula:

In this case, R ¹¹ and R ¹² independently represent hydrogen or hydrocarbyl, for example alkyl, or ^{the presence of R 11 together with R 12} and intervening atom(s) is a hetero having 4 to 8 atoms in the ring structure. complete the cycle

The term “protecting group” refers to a group of atoms that, when attached to a reactive functional group of a molecule, masks, reduces or prevents the reactivity of the functional group. Typically, protecting groups can be selectively removed as desired during the course of the synthesis. Examples of protecting groups are described in Greene and Wuts, Protective Groups in Organic Chemistry, 3 ^rd Ed., 1999, John Wiley & Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods , Vols. 1-8, 1971-1996, John Wiley & Sons, NY]. Representative nitrogen protecting groups are formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl (“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl -ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC") ), etc., but are not limited thereto. Representative hydroxyl protecting groups are those in which the hydroxyl group is acylated (esterified) or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (eg, TMS or TIPS). group), glycol ethers such as ethylene glycol and propylene glycol derivatives, and allyl ethers.

The present invention includes all pharmaceutically acceptable isotopically labeled compounds as described herein, wherein one or more atoms have the same number of atoms, but differ from an atomic mass or mass number generally found in nature. replaced by an atom having an atomic mass or mass number. In certain embodiments, the compounds of the present invention are enriched for such isotopically labeled substances (eg, compounds in which the isotopic distribution of the compound in the composition differs from the natural or typical isotopic distribution).

Examples of isotopes suitable for inclusion in the compounds of the present invention include isotopes of hydrogen such as ² H and ³ H, isotopes of carbon such as ¹¹ C, ¹³ C and ¹⁴ C, isotopes of chlorine such as, ³⁶ Cl, isotopes of fluorine such as ¹⁸ F, iodine such as ¹²³ I and ¹²⁵ I, isotopes of nitrogen such as ¹³ N and ¹⁵ N, isotopes of oxygen such as ¹⁵ O, ¹⁷ O and ¹⁸ O, isotopes of phosphorus such as ³² P, and isotopes of sulfur such as ³⁵ S.

Certain isotopically labeled compounds as disclosed herein, eg, those containing radioactive isotopes, are useful for drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, ie, ³ H and carbon-14, ie, ¹⁴ C, are useful for this purpose, given their ease of incorporation and preparation for detection.

Substitution with heavy isotopes such as deuterium, i.e. ² H, may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and thus, in certain circumstances may be preferred.

Substitution with positron emitting isotopes such as ¹¹ C, ¹⁸ F, ¹⁵ O and ¹³ N may be useful in positron emission tomography (PET) studies to examine substrate receptor occupancy.

In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched for one enantiomer. For example, a compound of the invention may contain greater than about 30% ee, about 40% ee, about 50% ee, about 60% ee, about 70% ee, about 80% ee, about 90% ee, or even about 95% ee. It can have more than % ee. In certain embodiments, compounds of the present invention may have more than one stereocenter. In certain such embodiments, the compounds of the present invention may be enriched for one or more diastereomers. For example, a compound of the invention may contain greater than about 30% de, about 40% de, about 50% de, about 60% de, about 70% de, about 80% de, about 90% de de, or even greater than about 95% de.

In certain embodiments, a therapeutic agent may be enriched to predominately provide one enantiomer of a compound (eg, of Formula I). An enantiomerically enriched mixture may comprise, for example, at least about 60 mole %, or more preferably at least about 75, about 90, about 95, or even about 99 mole % of one enantiomer. In certain embodiments, a compound enriched for one enantiomer is substantially free of the other enantiomer, where substantially free means that the substance is approximately free of the other enantiomer, e.g., in a composition or mixture of compounds, as compared to the amount of the other enantiomer. less than 10%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%. For example, if a composition or mixture of compounds contains about 98 g of a first enantiomer and about 2 g of a second enantiomer, it contains about 98 mole % of the first enantiomer and only about 2% of the second enantiomer. It is said to contain isomers.

In certain embodiments, the therapeutic agent may be enriched to provide predominantly one diastereomer of a compound (eg, of Formula I). A diastereomeric enriched mixture may comprise, for example, at least about 60 mole %, or more preferably at least about 75, about 90, about 95, or even about 99 mole % of one diastereomer.

The term "subject" contemplated for administration refers to a human (i.e., male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., a young man, middle-aged adults or the elderly)) and/or other primates (eg, cynomolgus monkeys, rhesus monkeys); mammals (including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs); and/or commercially relevant birds such as birds, including chickens, ducks, geese, quails, and/or turkeys. A preferred subject is a human.

As used herein, a therapeutic agent that "prevents" a disorder or condition reduces, in a statistical sample, the incidence of a disorder or condition in a treated sample compared to an untreated control sample, or is one of the disorder or condition compared to an untreated control sample. Refers to a compound that delays the onset of or reduces the severity of symptoms.

The term “treating” includes prophylactic and/or therapeutic treatment. The term “prophylactic or therapeutic” treatment is known in the art and includes administering to a subject one or more of the disclosed compositions. If it is administered prior to clinical manifestation of an unwanted condition (eg, a disease or other unwanted condition in a subject), the treatment is prophylactic (ie, it protects the subject against developing the unwanted condition); When it is administered after the manifestation of the unwanted condition, the treatment is therapeutic (ie, it is intended to reduce, alleviate or stabilize the pre-existing unwanted condition or its side effects).

The term “prodrug” is intended to include compounds that are converted under physiological conditions into a therapeutically active agent of the invention (eg, a compound of formula I). A common method of preparing prodrugs is to include one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the subject's enzymatic activity. For example, esters or carbonates (eg, esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compound of formula (I) in the formulations indicated above may be replaced, for example, by the corresponding suitable prodrug, wherein the hydroxyl of the parent compound is presented as an ester or carbonate or carboxylic acid do.

As used herein, an “effective amount” refers to an amount sufficient to achieve the desired biological effect. As used herein, “therapeutically effective amount” refers to an amount sufficient to achieve the desired therapeutic effect. For example, a therapeutically effective amount may refer to an amount sufficient to ameliorate at least one sign or symptom of cancer.

A "response" to a method of treatment is, among other things, reduction or amelioration of adverse symptoms, reduction of the progression of a disease or symptoms thereof, an increase in beneficial symptoms or clinical outcomes, attenuation of side effects, stabilization of the disease, partial or complete treatment of the disease. may include.

How to use

The PCSK9 gene was identified using gene mapping techniques for DNA of subjects with autosomal dominant hypercholesterolemia (Abifadel, et al. Nat. Genet. 2003 34:154-6). The encoded protein is a serine protease expressed primarily in the liver, intestine, kidney and nervous system. Without wishing to be bound by any particular theory, studies of gene mutations have shown that its putative role is in reducing LDL receptors on the cell surface independent of catalytic activity. (Abifadel, et al. Expert Opin. Ther. Pat. 2010 20:1547-71). Binding of PCSK9 to the receptor results in lysosomal degradation. This enhanced degradation increases the amount of circulating low-density lipoprotein LDL (LDL-c). PCSK9 is up-regulated by statins, SREBP-1a and SREBP-2, LXR agonists, and insulin, but not on dietary cholesterol, glucagon, ethinylestradiol, chenodeoxycholic acid and bile acid activated farnesoid X receptor (FXR). (Maxwell, et al. J. Lipid Res. 2003 44:2109-19; Person et al. Endocrinology 2009 150:1140-6; Langhi et al. FEBS Lett. 2008 582:949-55]). Since increased levels of PCSK9 decrease the amount of LDL receptors on the cell surface, a proportional LDL-cholesterol lowering result cannot be achieved by increasing the statin dose. Accordingly, disclosed herein are methods of treating a wide range of cardiovascular diseases and conditions that would benefit from lowering LDL-c by inhibiting PCSK9.

In certain embodiments, a method of inhibiting PCSK9 occurs in a subject in need thereof, thereby treating a disease or disorder mediated by PCSK9.

Also disclosed herein is a method of treating or preventing a disease or disorder mediated by PCSK9 comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof. In certain embodiments, disclosed herein is a method of treating a disease or disorder mediated by PCSK9 comprising administering a compound of Formula I, or a pharmaceutically acceptable salt thereof. In certain embodiments, disclosed herein is a method of preventing a disease or disorder mediated by PCSK9 comprising administering a compound of Formula I, or a pharmaceutically acceptable salt thereof. Prevention of cardiovascular events through inhibition of PCSK9 is described, for example, in Robinson et al., Artherosclerosis 2015 243:593-597.

Exemplary cardiovascular diseases and conditions include dyslipidemia, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hypoalphalipoproteinemia, metabolic syndrome, diabetic complications, atherosclerosis, stroke, vascular dementia, chronic kidney disease, coronary heart disease, coronary artery disease, retinopathy, inflammation, thrombosis, peripheral vascular disease or congestive heart failure.

In certain embodiments, exemplary cardiovascular diseases and conditions are hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome and coronary artery disease. including, but not limited to. In certain embodiments, the disease is hypercholesterolemia, such as familial hypercholesterolemia or autosomal dominant hypercholesterolemia. In certain embodiments, the disease is hyperlipidemia. In certain embodiments, the disease is coronary artery disease.

In certain embodiments, the disclosed methods of treatment are capable of reducing high levels of circulating serum cholesterol, such as LDL-cholesterol and VLDL-cholesterol. The disclosed methods are also useful for reducing circulating serum triglycerides, circulating serum lipoprotein A, circulating serum LDL and atherogenic lipoproteins. In certain embodiments, the diseases or conditions treated with the disclosed compounds and compositions include atherosclerosis and atherosclerotic plaque formation. Subjects with gain-of-function mutations in the PCSK9 gene would also benefit from treatment with the disclosed compounds and compositions that counteract the mutation through inhibition of PCSK9.

Inhibition of PCSK9 has also shown therapeutic benefit in treating sepsis in a subject. Septic shock is an often fatal complication of a severe microbial infection (sepsis) that causes an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis occurs in microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharides (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors (Toll-like receptors (TLRs)) and therefore appears prominently in the septic inflammatory response (septic shock or sepsis). PCSK9 reduces LPS uptake by LDL receptors in the liver, overstimulating the body's immune response to pathogens in which free LPS causes sepsis. Inhibiting PCSK9 is beneficial in maintaining hepatic LDL receptors to achieve systemic pathogen clearance and detoxification for sepsis (see, e.g., Walley et al Sci. Translat. Med. 2014 6:1-10). .

Disclosed herein is a method of treating sepsis or septic shock comprising administering a compound of Formula I, or a pharmaceutically acceptable salt thereof. In certain embodiments, the disclosed methods of treatment are useful for increasing LPS absorption. Certain embodiments provide a method of reducing an inflammatory response induced by sepsis or septic shock.

combination therapy

The disclosed compounds and compositions may be administered in combination with other therapeutic agents, such as other agents suitable for the treatment of high levels of LDL and triglycerides. In certain embodiments, administering one or more additional therapeutic agents in combination with a compound of the present invention provides a synergistic effect. In certain embodiments, co-administering one or more additional therapeutic agents provides an additive effect.

In some embodiments, a method of treating a disease or disorder mediated by PCSK9 comprises concurrently administering a PCSK9 inhibitor with one or more other therapeutic agent(s). In some embodiments, the PCSK9 inhibitor is a compound of Formula (I). Other therapeutic agents may include monoclonal antibodies such as alirocumab, evolocumab, bococizumab, RG7652, LY3015014 and mAb316P. Additional exemplary antibodies are WO/2015/140079, WO/2015/142668, WO/2015/123423, WO/2015/073494, WO/2015/054619, WO/2014/197752, WO/2014/194111, WO/ 2014/194168, WO/2014/028354, WO/2013/039969, WO/2013/016648, WO/2012/101251, WO/2012/101253, WO/2012/101252, WO/2014/209384, WO/2014/ 150983, WO/2014/144080, WO/2013/166448, WO/2012/154999, WO/2013/008185, WO/2015/200438, WO/2014/107739, WO/2013/188855, WO/2013/169886, WO/2013/148284, WO/2013/091103, WO/2013/039958, WO/2012/177741, WO/2012/168491, WO/2012/170607, WO/2012/109530, WO/2012/088313, WO/ 2012/054438, WO/2011/072263, WO/2011/053759, WO/2011/053783, and WO/2011/037791. Other therapeutic agents that may be used in combination with the disclosed compounds and compositions include plant alkaloids and flavanoids known to have cholesterol lowering activity, such as berberine and quercetin. In some embodiments, small cholesterol lowering molecules such as ezetimbe and policosanol may be administered as additional therapeutic agents. In certain embodiments, polypeptides such as BMS-962476 and fibronectin-based scaffold domain proteins can be administered in combination with the disclosed compounds and compositions. Further exemplary compounds are WO/2014/150326, WO/2014/150395, WO/2014/139008, WO/2014/140210, WO/2014/127316, WO/2014/005224, WO/2013/177536, WO/ 2011/152508, WO/2011/130354, WO/2011/006000, and WO/2015/077154.

In certain embodiments, the additional therapeutic agent may be a statin such as atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin. In another embodiment, the disclosed compounds may be administered in combination with HMG-CoA reductase.

Any HMG-CoA reductase inhibitor may be used in the combination aspect of the present invention. The conversion of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) to mevalonate is an initial and rate limiting step in the cholesterol biosynthetic pathway. This step is catalyzed by the enzyme HMG-CoA reductase. Statins inhibit HMG-CoA reductase from catalyzing this conversion.

The term HMG-CoA reductase inhibitor refers to a compound that inhibits the bioconversion of hydroxymethylglutaryl-coenzyme A to mevalonic acid catalyzed by the enzyme HMG-CoA reductase. Such inhibition is readily determined by one skilled in the art according to standard assays (eg, Meth. Enzymol. 1981; 71:455-509 and references cited therein). Although a variety of such compounds are described and referenced below, other HMG-CoA reductase inhibitors will be known to those skilled in the art. U.S. Patent No. 4,231,938 (the disclosure of which is incorporated herein by reference) discloses certain compounds isolated after culturing of microorganisms belonging to the genus Aspergillus, such as rosuvastatin. Also, US Pat. No. 4,444,784, the disclosure of which is incorporated herein by reference, discloses synthetic derivatives of the aforementioned compounds, such as simvastatin. Also, US Pat. No. 4,739,073, the disclosure of which is incorporated by reference, discloses certain substituted indoles, such as fluvastatin. Also, US Pat. No. 4,346,227, the disclosure of which is incorporated by reference, discloses ML-236B derivatives such as pravastatin. Also, EP-491226A, the disclosure of which is incorporated by reference, discloses certain pyridyldihydroxyheptenoic acids such as cerivastatin. Also, U.S. Pat. No. 5,273,995, the disclosure of which is incorporated by reference, discloses certain 6-[2-(substituted-pyrrol-1-yl)alkyl]pyran-2-ones, such as atorvastatin, and any pharmaceutically acceptable thereof. Possible forms (ie, LIPITOR®) are disclosed. Additional HMG-CoA reductase inhibitors include rosuvastatin and pitavastatin.

Atorvastatin calcium (ie, atorvastatin hemicalcium) disclosed in US Pat. No. 5,273,995, which is incorporated herein by reference, is now marketed as Lipitor®.

Statins are also known as U.S. rosuvastatin disclosed in RE37,314 E, pitivastatin disclosed in EP 304063 BI and US 5,011,930, U.S. Pat. simvastatin disclosed in 4,444,784; U.S. Pat., which is incorporated herein by reference. 5,502,199; cerivastatin; U.S. Pat., which is incorporated herein by reference. 3,983,140; mevastatin; U.S. Pat., which is incorporated herein by reference. 4,448,784 and U.S. velostatin disclosed in No. 4,450,171; U.S. Pat., which is incorporated herein by reference. 4,739,073; fluvastatin; U.S. Pat., which is incorporated herein by reference. 4,804,770; U.S. Pat., which is incorporated herein by reference. 4,231,938; lovastatin; dalvastatin disclosed in European Patent Application Publication No. 738510 A2; fluindostatin disclosed in European Patent Application Publication No. 363934 Al; and U.S. Pat. 4,450,171, such as dihydrocompactin.

Any HMG-CoA synthetase inhibitor may be used in the combination embodiment of the present invention. The term HMG-CoA synthetase inhibitor refers to a compound that inhibits the biosynthesis of acetyl-coenzyme A and acetoacetyl-coenzyme A to hydroxymethylglutaryl-coenzyme A catalyzed by the enzyme HMG-CoA synthetase. Such inhibition is readily determined by those skilled in the art according to standard assays (Meth Enzymol. 1975; 35:155-160: Meth. Enzymol. 1985; 110:19-26 and references cited therein). . Although a variety of such compounds are described and referenced below, other HMG-CoA synthetase inhibitors will be known to those skilled in the art. U.S. Patent No. 5,120,729, the disclosure of which is incorporated herein by reference, discloses certain beta-lactam derivatives. U.S. Pat. No. 5,064,856, the disclosure of which is incorporated herein by reference, discloses certain spiro-lactone derivatives prepared by culturing a microorganism (MF5253). U.S. Pat. No. 4,847,271, the disclosure of which is incorporated herein by reference, discloses certain oxetane compounds, such as 11-(3-hydroxymethyl-4-oxo-2-oxetile)-3,5,7-trimethyl- 2,4-undeca-dienoic acid derivatives are disclosed.

Any compound that reduces HMG-CoA reductase gene expression may be used in the combined aspect of the present invention. These agents may be HMG-CoA reductase transcription inhibitors that block transcription of DNA or translation inhibitors that prevent or reduce translation of mRNA encoding HMG-CoA reductase into proteins. Such compounds may directly affect transcription or translation, or may be converted in vivo to compounds having the aforementioned activities by one or more enzymes in the cholesterol biosynthetic cascade, or may inhibit the accumulation of isoprene metabolites having the aforementioned activities. can cause Such compounds can produce this effect by inhibiting the activity of site-1 proteases (S1P), agonizing oxysterol receptors, or antagonizing SCAP, thereby reducing the level of SREBP (sterol regulatory element binding protein). Such adjustments are readily determined by one skilled in the art according to standard assays (Meth. Enzymol. 1985; 110:9-19). Although several compounds are described and referenced below, other inhibitors of HMG-CoA reductase gene expression will be known to those skilled in the art. U.S. Patent No. 5,041,432, the disclosure of which is incorporated by reference, discloses certain 15-substituted lanosterol derivatives.

Other oxygenated sterols that inhibit the synthesis of HMG-CoA reductase are described in E.l. Mercer (Prog. Lip. Res. 1993;32:357-416).

Any compound having activity as a CETP inhibitor may act as a second compound in the combination therapy embodiment of the present invention. The term CETP inhibitor refers to compounds that inhibit cholesteryl ester transfer protein (CETP) mediated transport of various cholesteryl esters and triglycerides from HDL to LDL and VLDL. Such CETP inhibitory activity is readily determined by one of ordinary skill in the art according to standard assays (eg, US Pat. No. 6,140,343). Various CETP inhibitors will be known to those skilled in the art and are disclosed, for example, in commonly assigned US Pat. No. 6,140,343 and commonly assigned US Pat. No. 6,197,786. CETP inhibitors are also described in US Pat. No. 6,723,752, which is (2R)-3-{[3-(4-chloro-3-ethyl-phenoxy)-phenyl]-[[3-(1,1) ,2,2-tetrafluoro-ethoxy)-phenyl]-methyl]-amino}-1,1,1-trifluoro-2-propanol. In addition, CETP inhibitors incorporated herein are described in US Patent Application Serial No. 10/807838, filed March 23, 2004. U.S. Pat. No. 5,512,548 discloses certain polypeptide derivatives having activity as CETP inhibitors, while certain CETP inhibitory rosenolactone derivatives and phosphate-containing analogs of cholesteryl esters, respectively, are described in J. Antibiot, 49(8) 815-816 (1996) and Bioorg. Med. Chem. Lett.; 6:1951-1954 (1996)].

Any PPAR modulator may be used in the combination aspect of the present invention. The term PPAR modulator refers to a compound that modulates peroxisome proliferator activator receptor (PPAR) activity in mammals, particularly humans. Such adjustments are readily determined by those skilled in the art according to standard assays known in the literature. These compounds regulate the transcription of key genes involved in lipid and glucose metabolism by modulating the PPAR receptor, such as those involved in fatty acid oxidation and high-density lipoprotein (HDL) assembly (eg, apolipoprotein Al gene transcription) throughout the body. It is thought to reduce fat and increase HDL cholesterol. Because of their activity, these compounds increase HDL cholesterol and apolipoprotein A1 in mammals, particularly humans, while also reducing plasma levels of triglycerides, VLDL cholesterol, LDL cholesterol and their related components, such as apolipoprotein B. For this reason, these compounds are useful in the treatment and correction of a variety of dyslipidemias observed to be associated with the development and development of atherosclerosis and cardiovascular diseases (including hypoalphalipoproteinemia and hypertriglyceridemia). A variety of such compounds are described and referenced below, although others will be known to those skilled in the art. International Application Nos. WO 02/064549 and 02/064130 and U.S. Patent Application No. 10/720942, filed on November 24, 2003, and U.S. Patent Application Serial No. 60/552114, filed March 10, 2004, of The disclosure is incorporated herein by reference) discloses certain compounds that are PPARa activators.

Any other PPAR modulator may be used in the combination aspect of the present invention. In particular, modulators of PPARp and/or PPARy may be useful in combination with the compounds of the present invention. An example of a PPAR inhibitor is described in US 2003/0225158 as {5-Methoxy-2-methyl-4-[4-(4-trifluoromethyl-benzyloxy)-benzylsulfanyl]-phenoxy}-acetic acid have.

Any MTP/apo B (microsomal triglyceride transfer protein and/or apolipoprotein B) secretion inhibitor may be used in the combination aspect of the present invention. The term MTP/apo B secretion inhibitor refers to compounds that inhibit the secretion of triglycerides, cholesteryl esters and phospholipids. Such inhibition is readily determined by one skilled in the art according to standard assays (eg Wetterau, J. R. 1992; Science 258:999). Various inhibitors of MTP/apo B secretion will be known to those skilled in the art, including implitapide (Bayer) and additional compounds such as those disclosed in WO 96/40640 and WO 98/23593 (two exemplary publications).

Any squalene synthetase inhibitor may be used in the combination embodiment of the present invention. The term squalene synthase inhibitor refers to a compound that inhibits the condensation reaction of two molecules of farnesylpyrophosphate to form squalene catalyzed by the enzyme squalene synthase. Such inhibition is readily determined by one of ordinary skill in the art according to standard assays (Meth. Enzymol. 1969; 15: 393-454 and Meth. Enzymol. 1985; 110:359-373 and references incorporated therein). do. Although a variety of such compounds are described and referenced below, other squalene synthase inhibitors will be known to those skilled in the art. U.S. Patent No. 5,026,554, the disclosure of which is incorporated by reference, discloses fermentation products of microorganism MF5465 (ATCC 74011), including zaragozic acid. A summary of other patented squalene synthase inhibitors has been compiled (Curr. Op. Then Patents (1993) 861-4).

Any squalene epoxidase inhibitor may be used in the combined embodiment of the present invention. The term squalene epoxidase inhibitor refers to a compound that inhibits the bioconversion of squalene and molecular oxygen to squalene-2,3-epoxide catalyzed by the enzyme squalene epoxidase. Such inhibition is readily determined by one skilled in the art according to standard assays (Biochim. Biophys. Acta 1984; 794:466-471). Although a variety of such compounds are described and referenced below, other squalene epoxidase inhibitors will be known to those skilled in the art. U.S. Patent Nos. 5,011,859 and 5,064,864, the disclosures of which are incorporated by reference, disclose certain fluoro analogs of squalene. EP Publication No. 395,768 A, the disclosure of which is incorporated by reference, discloses certain substituted allylamine derivatives. PCT Publication No. WO 9312069 A, the disclosure of which is incorporated herein by reference, discloses certain amino alcohol derivatives. U.S. Patent No. 5,051,534, the disclosure of which is incorporated herein by reference, discloses certain cyclopropyloxy-squalene derivatives.

Any squalene cyclase inhibitor may be used in the combination embodiment of the present invention as the second component. The term squalene cyclase inhibitor refers to a compound that inhibits the bioconversion of squalene-2,3-epoxide to lanosterol catalyzed by the enzyme squalene cyclase. Such inhibition is readily determined by one skilled in the art according to standard assays (FEBS Lett. 1989; 244:347-350). Also, while the compounds described and referenced below are squalene cyclase inhibitors, other squalene cyclase inhibitors will be known to those skilled in the art. Pd Publication No. WO9410150, the disclosure of which is incorporated herein by reference, describes certain 1,2,3,5,6,7,8,8a-octahydro-5,5,8(beta)-trimethyl-6- Isoquinolinamine derivatives such as N-trifluoroacetyl-1,2,3,5,6,7,8,8a-octahydro-2-allyl-5,5,8(beta)-trimethyl-6(beta) )-isoquinolinamines. French Patent Publication No. 2697250, the disclosure of which is incorporated herein by reference, discloses certain beta, beta-dimethyl-4-piperidine ethanol derivatives such as 1-(1,5,9-trimethyldecyl)-beta,beta -Dimethyl-4-piperidineethanol is disclosed.

Any combined squalene epoxidase/squalene cyclase inhibitor may be used in the combined embodiment of the present invention as the second component. The term combined squalene epoxidase/squalene cyclase inhibitor refers to a compound that inhibits the bioconversion of squalene to lanosterol via the squalene-2,3-epoxide intermediate. In some assays, the distinction between squalene epoxidase inhibitors and squalene cyclase inhibitors is not possible; However, such assays are recognized by those skilled in the art. Therefore, inhibition by a combined squalene epoxidase/squalene cyclase inhibitor is readily determined by one skilled in the art according to the standard assays described above for squalene cyclase or squalene epoxidase inhibitors. Although a variety of such compounds are described and referenced below, other squalene epoxidase/squalene cyclase inhibitors will be known to those skilled in the art. U.S. Pat. Nos. 5,084,461 and 5,278,171, the disclosures of which are incorporated by reference, disclose certain azadecalin derivatives. EP Publication No. 468,434, the disclosure of which is incorporated by reference, discloses certain piperidyl ethers and thio-ether derivatives such as 2-(1-piperidyl)pentyl isopentyl sulfoxide and 2-(1-piperidyl). ) ethyl ethyl sulfide. PCT Publication No. WO 9401404, the disclosure of which is incorporated herein by reference, discloses certain acyl-piperidines such as 1-(1-oxopentyl-5-phenylthio)-4-(2-hydroxy-1- methyl)-ethyl)piperidine. U.S. Pat. No. 5,102,915, the disclosure of which is incorporated herein by reference, discloses certain cyclopropyloxy-squalene derivatives.

The compounds of the present invention may also be administered in combination with naturally occurring compounds that act to lower plasma cholesterol levels. These naturally occurring compounds are commonly referred to as nutraceuticals and include, for example, garlic extract and niacin. A sustained release form of niacin is available and is known as Niaspan. In addition, niacin may be combined with other therapeutic agents, such as lovastatin, or another HMG-CoA reductase inhibitor. This combination therapy with lovastatin is known as ADVICOR™ (Kos Pharmaceuticals Inc.).

Any cholesterol absorption inhibitor may be used in the combination embodiment of the present invention as an additional compound. The term cholesterol absorption inhibition refers to the ability of a compound to prevent cholesterol contained within the intestinal lumen from entering and/or passing from within the intestinal cells into the lymphatic system and/or the bloodstream. Such cholesterol absorption inhibitory activity is readily determined by those skilled in the art according to standard assays (eg, J. Lipid Res. (1993) 34: 377-395). Cholesterol absorption inhibitors are known to the person skilled in the art and are described, for example, in PCT WO 94/00480. An example of a cholesterol absorption inhibitor is ZETIA™ (ezetimibe) (Schering-Plow/Merck).

Any ACAT inhibitor may be used in the combination therapy aspect of the present invention. The term ACAT inhibitor refers to a compound that inhibits the intracellular esterification of dietary cholesterol by the enzyme acyl CoA:cholesterol acyltransferase. Such inhibition can be achieved by standard assays such as those described in Heider et al. Journal of Lipid Research., 24:1127 (1983)]. A variety of such compounds are known to those skilled in the art, for example, US Pat. No. 5,510,379 discloses certain carboxysulfonates, while WO 96/26948 and WO 96/10559 disclose urea derivatives with ACAT inhibitory activity. Examples of ACAT inhibitors include compounds such as Abashimibe (Pfizer), CS-505 (Sankyo) and Eflusimibe (Eli Lilly and Pierre Fabre).

Lipase inhibitors may be used in combination therapy embodiments of the present invention. Lipase inhibitors are compounds that inhibit the metabolic cleavage of dietary triglycerides or plasma phospholipids into free fatty acids and the corresponding glycerides (eg EL, HL, etc.). Under normal physiological conditions, lipolysis occurs through a two-step process involving acylation of the activated serine residue of the lipase enzyme. This results in the production of a fatty acid-lipase hemiacetal intermediate, which is then cleaved to release diglycerides. After further deacylation, the lipase-fatty acid intermediate is cleaved, resulting in free lipase, glycerides and fatty acids. In the intestine, the free fatty acids and monoglycerides produced are incorporated into bile acid-phospholipid micelles, which are subsequently absorbed at the level of the brush border of the small intestine. The micelles eventually enter the peripheral circulation as chylomicrons. Such lipase inhibitory activity is readily determined by one skilled in the art according to standard assays (eg Methods Enzymol. 286: 190-231).

Pancreatic lipase mediates the metabolic cleavage of fatty acids from triglycerides at the 1- and 3-carbon positions. The primary sites of ingested fat metabolism are in the duodenum and proximal jejunum, usually by pancreatic lipase, which is secreted in excess for the breakdown of fat in the upper small intestine. Because pancreatic lipase is a key enzyme required for absorption of dietary triglycerides, inhibitors have utility in the treatment of obesity and other related conditions. Such pancreatic lipase inhibitory activity is readily determined by those skilled in the art according to standard assays (eg, Methods Enzymol. 286: 190-231).

Gastric lipases are immunologically distinct lipases responsible for the digestion of about 10 to 40% of dietary fat. Gastric lipases are secreted in response to mechanical stimulation, food intake, the presence of fatty foods, or by the sympathetic nervous system. Gastric lipolysis of ingested fat is physiologically important for the supply of fatty acids necessary to trigger pancreatic lipase activity in the intestine, and is also important for fat absorption in various physiological and pathological conditions related to pancreatic insufficiency. For example, C.K. Abrams, et al., Gastroenterology, 92,125 (1987)]. Such gastric lipase inhibitory activity is readily determined by those skilled in the art according to standard assays (eg, Methods Enzymol. 286: 190-231).

Various gastric lipase inhibitors and/or pancreatic lipase inhibitors are known to those skilled in the art. Preferred lipase inhibitors are selected from lipstatin, tetrahydrolipstatin (orlistat), valilactone, esterastin, everlactone A and everlactone B. The compound tetrahydrolipstatin is particularly preferred. The lipase inhibitor N-3-trifluoromethylphenyl-N'-3-chloro-4'-trifluoromethylphenylurea and various urea derivatives related thereto are disclosed in US Pat. No. 4,405,644. Esteraxin, a lipase inhibitor, is disclosed in US Pat. Nos. 4,189,438 and 4,242,453. The lipase inhibitor cyclo-O,O'-[(1,6-hexanediyl)-bis-(iminocarbonyl)]dioxime and its various bis(iminocarbonyl)dioximes are described in Petersen et al. ., Liebig's Annalen, 562, 205-229 (1949).

Various pancreatic lipase inhibitors are described herein below. Pancreatic lipase inhibitor Lipstatin, (2S, 3S, 5S, 7Z, 10Z)-5-[(S)-2-formamido-4-methyl-valeryloxy]-2-hexyl-3-hydroxy- 7,10-hexadecanoic acid lactone, and tetrahydrolipstatin (orlistat), (2S, 3S, 5S)-5-[(S)-2-formamido-4-methyl-valeryloxy]-2 -Hexyl-3-hydroxy-hexadecanoic 1,3 acid lactones, and variously substituted N-formylucine derivatives and stereoisomers thereof, are disclosed in US Pat. No. 4,598,089. For example, tetrahydrolipstatin is disclosed, for example, in U.S. Patent Nos. 5,274,143; 5,420,305; 5,540,917; and 5,643,874. A pancreatic lipase inhibitor, FL-386, 1-[4-(2-methylpropyl)cyclohexyl]-2-[(phenylsulfonyl)oxy]-ethanone, and various substituted sulfonate derivatives related thereto Patent No. 4,452,813. Pancreatic lipase inhibitors, WAY-121898, 4-phenoxyphenyl-4-methylpiperidin-1-yl-carboxylate, and various carbamate esters and pharmaceutically acceptable salts related thereto are disclosed in U.S. Patent Nos. 5,512,565; 5,391,571; and 5,602,151. A pancreatic lipase inhibitor, valilactone, and a method for its preparation by microbial culture of the actinomycetes strain MG147-CF2 are disclosed in Kitahara, et al., J. Antibiotics, 40 (11), 1647-1650 (1987). has been Pancreatic lipase inhibitors, everactone A and everactone B, and a method for their preparation by microbial culture of the actinomycetes strain MG7-G1 are described in Umezawa, et al., J. Antibiotics, 33, 1594-1596 (1980). is disclosed in The use of everlactones A and B in inhibiting monoglyceride formation is disclosed in Japanese Patent Application Laid-Open No. 08-143457 published on Jun. 4, 1996.

Other compounds marketed for hyperlipidemia, including hypercholesterolemia, and intended to aid in the prevention or treatment of atherosclerosis, include bile acid sequestrants such as Welchol®, Colestid®, LoCholest ®) and Questran®; and fibric acid derivatives such as Atromid®, Lopid® and Tricor®.

Given the link between diabetes and atherosclerosis (eg metabolic syndrome), the compound of formula (I) may be administered in combination with an antidiabetic compound. Diabetes mellitus can be caused by combining a therapeutically effective amount of a compound of the invention with other agents (e.g. insulin) that can be used to treat diabetes, such as diabetes (especially type II), insulin resistance, impaired glucose tolerance, metabolic syndrome, etc. or any diabetic complications such as neuropathy, nephropathy, retinopathy or cataracts. This includes the class of antidiabetic agents (and certain agents) described herein.

Any glycogen phosphorylase inhibitor may be used as the second agent in combination with the compounds of the present invention. The term glycogen phosphorylase inhibitor refers to a compound that inhibits the bioconversion of glycogen to glucose-1-phosphate catalyzed by the enzyme glycogen phosphorylase. Such glycogen phosphorylase inhibitory activity is readily determined by those skilled in the art according to standard assays (eg, J. Med. Chem. 41 (1998) 2934-2938). A variety of glycogen phosphorylase inhibitors are known to those skilled in the art, including those described in WO 96/39384 and WO 96/39385.

Any aldose reductase inhibitor may be used in combination with the compounds of the present invention. The term aldose reductase inhibitor refers to a compound that inhibits the bioconversion of glucose to sorbitol catalyzed by the enzyme aldose reductase. Aldose reductase inhibition is readily determined by one skilled in the art according to standard assays (eg, J. Malone, Diabetes, 29:861-864 (1980). "Red Cell Sorbitol, an Indicator of Diabetic Control"). . A variety of aldose reductase inhibitors are known to those skilled in the art and are described in, for example, US Pat. No. 6,579,879, including 6-(5-chloro-3-methyl-benzofuran-2-sulfonyl)-2H-pyridazin-3-one. it has been described

Any sorbitol dehydrogenase inhibitor may be used in combination with the compounds of the present invention. The term sorbitol dehydrogenase inhibitor refers to a compound that inhibits the bioconversion of sorbitol to fructose catalyzed by the enzyme sorbitol dehydrogenase. Such sorbitol dehydrogenase inhibitor activity is readily determined by those skilled in the art according to standard assays (eg Analyt. Biochem (2000) 280: 329-331). Various sorbitol dehydrogenase inhibitors are known, for example, US Pat. Nos. 5,728,704 and 5,866,578 disclose compounds and methods of treating or preventing diabetic complications by inhibiting the enzyme sorbitol dehydrogenase.

Any glucosidase inhibitor may be used in combination with the compounds of the present invention. Glucosidase inhibitors inhibit the enzymatic hydrolysis of complex carbohydrates to bioavailable monosaccharides such as glucose by glycoside hydrolases such as amylase or maltase. The rapid metabolic action of glucosidase, especially after ingestion of high levels of carbohydrates, leads to a state of dietary hyperglycemia, which in adipose tissue or diabetic subjects leads to enhanced secretion of insulin, increased fat synthesis and reduction of lipolysis. cause This hyperglycemia is often followed by hypoglycemia due to the increased levels of insulin present. It is also known that the yumi remaining in the stomach promotes the production of gastric juice, which initiates or promotes the development of gastritis or duodenal ulcer. Thus, glucosidase inhibitors are known to be useful for accelerating the passage of carbohydrates through the stomach and inhibiting the absorption of glucose from the intestine. In addition, the conversion of carbohydrates into adipose tissue into lipids and subsequent incorporation of dietary fat into adipose tissue precipitates is reduced or delayed, with the concomitant benefit of reducing or preventing harmful abnormalities resulting therefrom. Such glucosidase inhibitory activity is readily determined by those skilled in the art according to standard assays (eg, Biochemistry (1969) 8: 4214).

Generally preferred glucosidase inhibitors include amylase inhibitors. Amylase inhibitors are glucosidase inhibitors that inhibit the enzymatic breakdown of starch or glycogen to maltose. Such amylase inhibitory activity is readily determined by those skilled in the art according to standard assays (eg Methods Enzymol. (1955) 1:149). Inhibition of this enzymatic degradation reduces the amount of bioavailable sugars including glucose and maltose and is beneficial in the concomitant deleterious conditions resulting therefrom.

Various glucosidase inhibitors are known to those skilled in the art, examples are provided below. Preferred glucosidase inhibitors are selected from acarbose, adiposin, voglibose, miglitol, emiglitate, camiglibose, tendamistate, trestatin, pradimycin-Q and salvostatin. Acarbose, a glucosidase inhibitor, and various amino sugar derivatives related thereto are disclosed in US Pat. Nos. 4,062,950 and 4,174,439, respectively. Adiposin, a glucosidase inhibitor, is disclosed in US Pat. No. 4,254,256. Glucosidase inhibitor voglibose, 3,4-dideoxy-4-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-2-C-(hydroxymethyl)-D-epi- inositol), and various N-substituted pseudo-amino sugars related thereto, are disclosed in US Pat. No. 4,701,559. miglitol, a glucosidase inhibitor, (2R,3R,4R,5S)-1-(2-hydroxyethyl)-2-(hydroxymethyl)-3,4,5-piperidinetriol), and Various 3,4,5-trihydroxypiperidines related thereto are disclosed in US Pat. No. 4,639,436. glucosidase inhibitor emiglitate, ethyl p-[2-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]ethoxy] -benzoate), and various derivatives related thereto and pharmaceutically acceptable acid addition salts thereof, are disclosed in US Pat. No. 5,192,772. glucosidase inhibitor MDL-25637, 2,6-dideoxy-7-O-β-D-glucopyrano-syl-2,6-imino-D-glycero-L-gluco-heptitol) , various homodisaccharides related thereto and pharmaceutically acceptable acid addition salts thereof are disclosed in US Pat. No. 4,634,765. Glucosidase inhibitor camiglibose, methyl 6-deoxy-6-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]- aD-glucopyranoside sesquihydrate, deoxy-nojirimycin derivatives related thereto, various pharmaceutically acceptable salts thereof, and synthetic methods for their preparation are disclosed in US Pat. Nos. 5,157,116 and 5,504,078. The glucosidase inhibitor salvostatin and various pseudosaccharides related thereto are disclosed in US Pat. No. 5,091,524.

A variety of amylase inhibitors are known to those skilled in the art. The amylase inhibitor tendamistat and various cyclic peptides related thereto are disclosed in US Pat. No. 4,451,455. The amylase inhibitor AI-3688 and various cyclic polypeptides related thereto are disclosed in US Pat. No. 4,623,714. The amylase inhibitor trestatin (composed of a mixture of trestatin A, trestatin B and trestatin C) and various trehalose-containing amino sugars related thereto are disclosed in US Pat. No. 4,273,765.

Additional antidiabetic compounds that can be used as a second agent in combination with a compound of the present invention include, for example: biguanides (eg metformin), insulin secretagogues (eg sulfonylureas and glycans) need), glitazone, non-glitazone PPARy agonist, PPARp agonist, inhibitor of DPP-IV, inhibitor of PDE5, inhibitor of GSK-3, glucagon antagonist, inhibitor of f-1,6-BPase (Metabasis/ Sankyo), GLP-1/analog (AC 2993, also known as exendin-4), insulin and insulin mimetics (Merck natural products). Other examples include PKC-P inhibitors and AGE disruptors.

In addition, the compounds of the present invention may be used in combination with cardiovascular agents, such as antihypertensive agents. Any antihypertensive agent can be used as the second agent in such combination, examples are provided herein. Such antihypertensive activity is readily determined by one skilled in the art according to standard assays (eg blood pressure measurement).

Amlodipine and related dihydropyridine compounds are disclosed in US Pat. No. 4,572,909, incorporated herein by reference as potent antiischemic and antihypertensive agents. U.S. Patent No. 4,879,303, incorporated herein by reference, discloses amlodipine benzenesulfonate salt (also referred to as amlodipine besylate). Amlodipine and amlodipine besylate are strong and long-acting calcium channel blockers. As such, amlodipine, amlodipine besylate, amlodipine maleate and other pharmaceutically acceptable acid addition salts of amlodipine are useful as antihypertensive and antiischemic agents. Amlodipine besylate is currently marketed as Norvasc®.

Calcium channel blockers within the scope of this invention include, but are not limited to: bepridil (which may be prepared as disclosed in U.S. Patent No. 3,962,238 or U.S. Reissue Patent No. 30,577); kletiazem (which may be prepared as disclosed in U.S. Patent No. 4,567,175); diltiazem (which may be prepared as disclosed in U.S. Patent No. 3,562); pendiline (which may be prepared as disclosed in U.S. Patent No. 3,262,977); gallopamil (which may be prepared as disclosed in U.S. Patent No. 3,261,859); mibefradil (which may be prepared as disclosed in U.S. Patent No. 4,808,605); prenylamine (which may be prepared as disclosed in U.S. Patent No. 3,152,173); cemotiadil (which may be prepared as disclosed in U.S. Patent No. 4,786,635); terodilin (which may be prepared as disclosed in U.S. Patent No. 3,371,014); verapamil (which may be prepared as disclosed in U.S. Patent No. 3,261,859); aranifine (which may be prepared as disclosed in U.S. Patent No. 4,572,909); vanidipine (which may be prepared as disclosed in U.S. Patent No. 4,220,649); benidipine (which may be prepared as disclosed in European Patent Application Publication No. 106,275); clinidipine (which may be prepared as disclosed in U.S. Patent No. 4,672,068); eponidipine (which may be prepared as disclosed in U.S. Patent No. 4,885,284); ergodipine (which may be prepared as disclosed in U.S. Patent No. 4,952,592); felodipine (which may be prepared as disclosed in U.S. Patent No. 4,264,611); isradipine (which may be prepared as disclosed in U.S. Patent No. 4,466,972); racidipine (which may be prepared as disclosed in U.S. Patent No. 4,801,599); lecanidipine (which may be prepared as disclosed in U.S. Patent No. 4,705,797); manidipine (which may be prepared as disclosed in U.S. Patent No. 4,892,875); nicardipine (which may be prepared as disclosed in U.S. Patent No. 3,985,758); Nifedipine (which may be prepared as disclosed in U.S. Patent No. 3,485,847; Nilvadipine (may be prepared as disclosed in U.S. Patent No. 4,338,322); Nimodipine (may be prepared as disclosed in U.S. Patent No. 3,799,934) Yes; nisoldipine (which may be prepared as disclosed in US Pat. No. 4,154,839); nitrendipine (which may be prepared as disclosed in US Pat. No. 3,799,934); cinnarizine (which may be prepared as disclosed in US Pat. No. 2,882,271) Flunarizine (which may be prepared as disclosed in U.S. Patent No. 3,773,939; Lidoplazine (may be prepared as disclosed in U.S. Patent No. 3,267,104); Lomerizine (may be prepared as disclosed in U.S. Patent No. 3,267,104); 4,663,325); bencyclane (which may be prepared as disclosed in Hungarian Patent No. 151,865); etapanone (which may be prepared as disclosed in German Patent No. 1,265,758); and Perhexillin (can be prepared as disclosed in British Patent No. 1,025,578).The disclosure of all these U.S. patents is hereby incorporated by reference.Examples of currently marketed products containing antihypertensive agents include calcium channel blockers, such as Cardizem®, Adalat®, Calan®, Cardene®, Covera®, Dilacor®, DynaCirc®, Procadia X L (Procardia XL®), Sular®, Tiazac®, Vascor®, Verelan®, Isoptin®, Nimotop®, Norvasc ®) and Plendil®, angiotensin converting enzyme (ACE) inhibitors such as Accupril®, Altace®, Captopril®, Lotensin®, Mavic ( Mavik®), Monopril®, Prinivil® , Univasc®, Vasotec® and Zestril®.

Angiotensin converting enzyme inhibitors (ACE inhibitors) that are within the scope of the present invention include, but are not limited to: alacepril (which may be prepared as disclosed in US Pat. No. 4,248,883); benazepril (which may be prepared as disclosed in U.S. Patent No. 4,410,520); captopril (which may be prepared as disclosed in U.S. Patent Nos. 4,046,889 and 4,105,776); Ceronapril (which may be prepared as disclosed in U.S. Patent No. 4,452,790); delapril (which may be prepared as disclosed in U.S. Patent No. 4,385,051); enalapril (which may be prepared as disclosed in U.S. Patent No. 4,374,829); fosinopril (which may be prepared as disclosed in U.S. Patent No. 4,337,201); imadapril (which may be prepared as disclosed in U.S. Patent No. 4,508,727); lisinopril (which may be prepared as disclosed in U.S. Patent No. 4,555,502); mobeltopril (which may be prepared as disclosed in Belgian Patent No. 893,553); perindopril (which may be prepared as disclosed in U.S. Patent No. 4,508,729); quinapril (which may be prepared as disclosed in U.S. Patent No. 4,344,949); ramipril (which may be prepared as disclosed in U.S. Patent No. 4,587,258); Spirapril (which may be prepared as disclosed in U.S. Patent No. 4,470,972); Temocapril (may be prepared as disclosed in U.S. Patent No. 4,699,905); and trandolapril (which may be prepared as disclosed in U.S. Patent No. 4,933,361). The disclosures of all such US patents are incorporated herein by reference.

Angiotensin-II receptor antagonists (A-II antagonists) within the scope of the present invention include, but are not limited to: candesartan (which may be prepared as disclosed in U.S. Patent No. 5,196,444); eprosartan (which may be prepared as disclosed in U.S. Patent No. 5,185,351); ilbesartan (which may be prepared as disclosed in US Pat. No. 5,270,317); losartan (which may be prepared as disclosed in U.S. Patent No. 5,138,069); and valsartan (which may be prepared as disclosed in U.S. Patent No. 5,399,578). The disclosures of all such US patents are incorporated herein by reference.

Beta-adrenergic receptor blockers (beta- or β-blockers) within the scope of the present invention include, but are not limited to: acebutolol (which may be prepared as disclosed in U.S. Patent No. 3,857,952); alprenolol (which may be prepared as disclosed in Dutch Patent No. 6,605,692); amosularol (which may be prepared as disclosed in U.S. Patent No. 4,217,305); arotinolol (which may be prepared as disclosed in U.S. Patent No. 3,932,400); atenolol (which may be prepared as disclosed in U.S. Patent Nos. 3,663,607 or 3,836,671); befunolol (which may be prepared as disclosed in U.S. Patent No. 3,853,923); betaxolol (which may be prepared as disclosed in U.S. Patent No. 4,252,984); bevantolol (which may be prepared as disclosed in U.S. Patent No. 3,857,981); bisoprolol (which may be prepared as disclosed in U.S. Patent No. 4,171,370); bopindolol (which may be prepared as disclosed in U.S. Patent No. 4,340,541); bucumolol (which may be prepared as disclosed in U.S. Patent No. 3,663,570); bufetolol (which may be prepared as disclosed in U.S. Patent No. 3,723,476); bufuralol (which may be prepared as disclosed in U.S. Patent No. 3,929,836); bunitrrolol (which may be prepared as disclosed in U.S. Patent Nos. 3,940,489 and 3,961,071); buprandolol (which may be prepared as disclosed in U.S. Patent No. 3,309,406); butyridine hydrochloride (which may be prepared as disclosed in French Patent No. 1,390,056); butopyrrolol (which may be prepared as disclosed in U.S. Patent No. 4,252,825); carazolol (which may be prepared as disclosed in German Patent No. 2,240,599); cateolol (which may be prepared as disclosed in U.S. Patent No. 3,910,924); carvedilol (which may be prepared as disclosed in U.S. Patent No. 4,503,067); seliprolol (which may be prepared as disclosed in U.S. Patent No. 4,034,009); cetamolol (which may be prepared as disclosed in U.S. Patent No. 4,059,622); chloranolol (which may be prepared as disclosed in German Patent No. 2,213,044); dilevarol (which may be prepared as disclosed in Clifton et al., Journal of Medicinal Chemistry, 1982, 25, 670); epanolol (which may be prepared as disclosed in European Patent Application Publication No. 41,491); indenolol (which may be prepared as disclosed in U.S. Patent No. 4,045,482); labetalol (which may be prepared as disclosed in U.S. Patent No. 4,012,444); Levobunolol (which may be prepared as disclosed in U.S. Patent No. 4,463,176); mepindolol (which may be prepared as disclosed in Seeman et al., Helv. Chim. Acta, Chim. Acta, 1971, 54, 241); metipranolol (which may be prepared as disclosed in Czechoslovak Patent Application No. 128,471); metoprolol (which may be prepared as disclosed in U.S. Patent No. 3,873,600); morphrolol (which may be prepared as disclosed in U.S. Patent No. 3,501,7691); nadolol (which may be prepared as disclosed in U.S. Patent No. 3,935,267); nadoxolol (which may be prepared as disclosed in U.S. Patent No. 3,819,702); nebivalol (which may be prepared as disclosed in U.S. Patent No. 4,654,362); nipradilol (which may be prepared as disclosed in U.S. Patent No. 4,394,382); oxprenolol (which may be prepared as disclosed in British Patent No. 1,077,603); perbutolol (which may be prepared as disclosed in U.S. Patent No. 3,551,493); pindolol (which may be prepared as disclosed in Swiss Patent Nos. 469,002 and 472,404); fructolol (which may be prepared as disclosed in U.S. Patent No. 3,408,387); pronetalol (which may be prepared as disclosed in British Patent No. 909,357); propranolol (which may be prepared as disclosed in U.S. Patent Nos. 3,337,628 and 3,520,919); sotalol (which may be prepared as disclosed in Uloth et al., Journal of Medicinal Chemistry, 1966, 9, 88); sulfinalol (which may be prepared as disclosed in German Patent No. 2,728,641); Tallindole (which may be prepared as disclosed in U.S. Patent Nos. 3,935,259 and 4,038,313); tertatorol (which may be prepared as disclosed in U.S. Patent No. 3,960,891); tilisolol (which may be prepared as disclosed in U.S. Patent No. 4,129,565); timolol (which may be prepared as disclosed in U.S. Patent No. 3,655,663); toliprolol (which may be prepared as disclosed in US Pat. No. 3,432,545); and gibenolol (which may be prepared as disclosed in U.S. Patent No. 4,018,824). The disclosures of all such US patents are incorporated herein by reference.

Alpha-adrenergic receptor blockers (alpha- or a-blockers) within the scope of the present invention include, but are not limited to: amosulalol (which may be prepared as disclosed in U.S. Patent No. 4,217,307); arotinolol (which may be prepared as disclosed in U.S. Patent No. 3,932,400); dapiprazole (which may be prepared as disclosed in U.S. Patent No. 4,252,721); doxazosin (which may be prepared as disclosed in U.S. Patent No. 4,188,390); fenspiride (which may be prepared as disclosed in U.S. Patent No. 3,399,192); indoramine (which may be prepared as disclosed in U.S. Patent No. 3,527,761); labetolol (which may be prepared as disclosed above); naphthopidil (which may be prepared as disclosed in U.S. Patent No. 3,997,666); nicergoline (which may be prepared as disclosed in U.S. Patent No. 3,228,943); prazosin (which may be prepared as disclosed in U.S. Patent No. 3,511,836); tamsulosin (which may be prepared as disclosed in U.S. Patent No. 4,703,063); tolazoline (which may be prepared as disclosed in U.S. Patent No. 2,161,938); trimazosin (which may be prepared as disclosed in U.S. Patent No. 3,669,968); and yohimbine (which may be isolated from natural sources according to methods well known to those skilled in the art). The disclosures of all such US patents are incorporated herein by reference.

As used herein, the term “vasodilator” is meant to include cerebrovasodilators, coronary vasodilators and peripheral vasodilators. Cerebrovasodilators within the scope of the present invention include, but are not limited to: bencyclane (which may be prepared as disclosed above); cinnarizine (which may be prepared as disclosed above); Citicoline (isolated from natural sources as disclosed in Kennedy et al., Journal of the American Chemical Society, 1955, 77, 250) or as disclosed in Kennedy, Journal of Biological Chemistry, 1956, 222, 185 can be synthesized together); cyclandelate (which may be prepared as disclosed in U.S. Patent No. 3,663,597); cyclonicates (which may be prepared as disclosed in German Patent No. 1,910,481); diisopropylamine dichloroacetate (which may be prepared as disclosed in British Patent No. 862,248); evernamonin (which may be prepared as disclosed in Hermann et al., Journal of the American Chemical Society, 1979, 101, 1540); Fasudil (which may be prepared as disclosed in U.S. Patent No. 4,678,783); phenoxedil (which may be prepared as disclosed in U.S. Patent No. 3,818,021); flunarizine (which may be prepared as disclosed in U.S. Patent No. 3,773,939); ibudilast (which may be prepared as disclosed in U.S. Patent No. 3,850,941); ifenprodil (which may be prepared as disclosed in U.S. Patent No. 3,509,164); lomerizine (which may be prepared as disclosed in U.S. Patent No. 4,663,325); napronil (which may be prepared as disclosed in U.S. Patent No. 3,334,096); nicametate (which may be prepared as disclosed in Blicke et al., Journal of the American Chemical Society, 1942, 64, 1722); nicergoline (which may be prepared as described above); nimodipine (which may be prepared as disclosed in U.S. Patent No. 3,799,934); papaverine (which may be prepared as reviewed in Goldberg, Chem. Prod. Chem. News, 1954, 17, 371); pentipylline (which may be prepared as disclosed in German Patent No. 860,217); Tinophedrine (which may be prepared as disclosed in U.S. Patent No. 3,563,997); vincarmine (which may be prepared as disclosed in U.S. Patent No. 3,770,724); Vinpocetine (which may be prepared as disclosed in U.S. Patent No. 4,035,750); and Biquidyl (which may be prepared as disclosed in U.S. Patent No. 2,500,444). The disclosures of all such US patents are incorporated herein by reference.

Coronary vasodilators within the scope of the present invention include, but are not limited to: amotrifen (which may be prepared as disclosed in U.S. Patent No. 3,010,965); bendazole (which may be prepared as disclosed in J. Chem. Soc. 1958, 2426); benfurodyl hemisuccinate (which may be prepared as disclosed in U.S. Patent No. 3,355,463); benziodarone (which may be prepared as disclosed in U.S. Patent No. 3,012,042); chloracizine (which may be prepared as disclosed in British Patent No. 740,932); chromona (which may be prepared as disclosed in U.S. Patent No. 3,282,938); clobenfural (which may be prepared as disclosed in British Patent No. 1,160,925); clonitrate (can be prepared from propanediol according to methods well known to those skilled in the art, see, for example, Annalen, 1870, 155, 165); chloricromene (which may be prepared as disclosed in U.S. Patent No. 4,452,811); Dillazeb (which may be prepared as disclosed in U.S. Patent No. 3,532,685); dipyridamole (which may be prepared as disclosed in British Patent No. 807,826); droprenylamine (which may be prepared as disclosed in German Patent No. 2,521,113); efloxate (which may be prepared as disclosed in British Patent Nos. 803,372 and 824,547); erythrityl tetranitrate (which may be prepared by nitration of erythritol according to methods well known to those skilled in the art); etaphenone (which may be prepared as disclosed in German Patent No. 1,265,758); pendiline (which may be prepared as disclosed in U.S. Patent No. 3,262,977); floredil (which may be prepared as disclosed in German Patent No. 2,020,464); Ganglefen (which may be prepared as disclosed in U.S.S.R. No. 115,905); hexetrol (which may be prepared as disclosed in U.S. Patent No. 2,357,985); hexobendin (which may be prepared as disclosed in U.S. Patent No. 3,267,103); itramine tosylate (which may be prepared as disclosed in Swedish Patent No. 168,308); Kelin (which may be prepared as disclosed in Baxter et al., Journal of the Chemical Society, 1949, S 30); lidoplazine (which may be prepared as disclosed in U.S. Patent No. 3,267,104); mannitol hexanitrate (which may be prepared by nitration of mannitol according to methods well known to those skilled in the art); Medibazine (which may be prepared as disclosed in U.S. Patent No. 3,119,826); nitroglycerin; pentaerythritol tetranitrate (which may be prepared by nitration of pentaerythritol according to methods well known to those skilled in the art); pentrinitrile (which may be prepared as disclosed in German Patent No. 638,422-3); perhexylline (which may be prepared as disclosed above); pimepilline (which may be prepared as disclosed in U.S. Patent No. 3,350,400); prenylamine (which may be prepared as disclosed in U.S. Patent No. 3,152,173); propatyl nitrate (which may be prepared as disclosed in French Patent No. 1,103,113); trapidyl (which may be prepared as disclosed in East German Patent No. 55,956); trichromyl (which may be prepared as disclosed in U.S. Patent No. 2,769,015); trimetazidine (which may be prepared as disclosed in U.S. Patent No. 3,262,852); trolnitrate phosphate (which may be prepared by nitration of triethanolamine followed by precipitation with phosphoric acid, according to methods well known to those skilled in the art); Visnadine (which may be prepared as disclosed in U.S. Patent Nos. 2,816,118 and 2,980,699). The disclosures of all such US patents are incorporated herein by reference.

Peripheral vasodilators within the scope of the present invention include, but are not limited to: aluminum nicotinate (which may be prepared as disclosed in US Pat. No. 2,970,082); barmethane (which may be prepared as disclosed in Corrigan et al., Journal of the American Chemical Society, 1945, 67, 1894); bencyclane (which may be prepared as disclosed above); betahistine (which may be prepared as disclosed in Walter et al.; Journal of the American Chemical Society, 1941, 63, 2771); bradykinin (which may be prepared as disclosed in Hamburg et al., Arch. Biochem. Biochem. Biophys., 1958, 76, 252); brovincarmine (which may be prepared as disclosed in U.S. Patent No. 4,146,643); bufeniod (which may be prepared as disclosed in U.S. Patent No. 3,542,870); buflomedil (which may be prepared as disclosed in U.S. Patent No. 3,895,030); butalamine (which may be prepared as disclosed in U.S. Patent No. 3,338,899); cetiedil (which may be prepared as disclosed in French Patent No. 1,460,571); cyclonicates (which may be prepared as disclosed in German Patent No. 1,910,481); cinephazide (which may be prepared as disclosed in Belgian Patent No. 730,345); cinnarizine (which may be prepared as disclosed above); cyclandelate (which may be prepared as disclosed above); diisopropylamine dichloroacetate (which may be prepared as disclosed above); eledoisin (which may be prepared as disclosed in British Patent No. 984,810); phenoxedil (which may be prepared as disclosed above); flunarizine (which may be prepared as disclosed above); hepronicate (which may be prepared as disclosed in U.S. Patent No. 3,384,642); ifenprodil (which may be prepared as disclosed above); Iloprost (which may be prepared as disclosed in U.S. Patent No. 4,692,464); inositol niacinate (which may be prepared as disclosed in Badgett et al., Journal of the American Chemical Society, 1947, 69, 2907); isoxsuprine (which may be prepared as disclosed in U.S. Patent No. 3,056,836); kallidine (which may be prepared as disclosed in Biochem. Biophys. Res. Commun., 1961, 6, 210); kallikrein (which may be prepared as disclosed in German Patent No. 1,102,973); moxisilite (which may be prepared as disclosed in German Patent No. 905,738); napronil (which may be prepared as disclosed above); nicametate (which may be prepared as disclosed above); nicergoline (which may be prepared as disclosed above); nicofuranose (which may be prepared as disclosed in Swiss Patent No. 366,523); nilidrine (which may be prepared as disclosed in U.S. Patent Nos. 2,661,372 and 2,661,373); pentipylline (which may be prepared as disclosed above); pentoxifylline (which may be prepared as disclosed in US Pat. No. 3,422,107); pyrivedil (which may be prepared as disclosed in U.S. Patent No. 3,299,067); prostaglandin E-i (which may be prepared by any of the methods mentioned in Merck Index, Twelfth Edition, Budaveri, Ed., New Jersey, 1996, p. 1353); sulloctidyl (which may be prepared as disclosed in German Patent No. 2,334,404); tolazoline (which may be prepared as disclosed in U.S. Patent No. 2,161,938); and xanthinol niacinate (which may be prepared as disclosed in German Patent No. 1,102,750 or Korbonits et al., Acta. Pharm. Hung., 1968, 38, 98). The disclosures of all such US patents are incorporated herein by reference.

The term "diuretic" within the scope of the present invention includes the diuretic benzothiadiazine derivative, the diuretic organomercury, the diuretic purine, the diuretic steroid, the diuretic sulfonamide derivative, the diuretic uracil and other diuretics such as amanozine (as disclosed in Austrian Patent No. 168,063). can be prepared); amiloride (which may be prepared as disclosed in Belgian Patent No. 639,386); abutin (which may be prepared as disclosed in Tschitschibabin, Annalen, 1930, 479, 303); chlorazanil (which may be prepared as disclosed in Austrian Patent No. 168,063); ethacrynic acid (which may be prepared as disclosed in U.S. Patent No. 3,255,241); etozolin (which may be prepared as disclosed in U.S. Patent No. 3,072,653); hydracarbazine (which may be prepared as disclosed in British Patent No. 856,409); isosorbide (which may be prepared as disclosed in U.S. Patent No. 3,160,641); mannitol; methochalcone (which may be prepared as disclosed in Freudenberg et al., Ber., 1957, 90, 957); muzolimine (which may be prepared as disclosed in U.S. Patent No. 4,018,890); perhexylline (which may be prepared as disclosed above); ticrinafen (which may be prepared as disclosed in U.S. Patent No. 3,758,506); triamterene (which may be prepared as disclosed in U.S. Patent No. 3,081,230); and urea. The disclosures of all such US patents are incorporated herein by reference.

Diuretic benzothiadiazine derivatives within the scope of the present invention include, but are not limited to: althiazide (which may be prepared as disclosed in British Patent No. 902,658); bendroflumethiazide (which may be prepared as disclosed in U.S. Patent No. 3,265,573); benzthiazide (McManus et al., 136th Am. Soc. Meeting (Atlantic City, September 1959), Abstract of papers, pp 13-0); benzylhydrochlorothiazide (which may be prepared as disclosed in U.S. Patent No. 3,108,097); butiazide (which may be prepared as disclosed in British Patent Nos. 861,367 and 885,078); chlorothiazide (which may be prepared as disclosed in U.S. Patent Nos. 2,809,194 and 2,937,169); chlorthalidone (which may be prepared as disclosed in U.S. Patent No. 3,055,904); cyclopenthiazide (which may be prepared as disclosed in Belgian Patent No. 587,225); cyclothiazide (which may be prepared as disclosed in Whitehead et al., Journal of Organic Chemistry, 1961, 26, 2814); epithiazide (which may be prepared as disclosed in U.S. Patent No. 3,009,911); ethiazide (which may be prepared as disclosed in British Patent No. 861,367); fenquizone (which may be prepared as disclosed in U.S. Patent No. 3,870,720); Indapamide (which may be prepared as disclosed in US Pat. No. 3,565,911; hydrochlorothiazide, which may be prepared as disclosed in US Pat. No. 3,164,588); hydroflumethiazide (which may be prepared as disclosed in US Pat. No. 3,254,076) Meticlotiazide (which may be prepared as disclosed in Close et al., Journal of the American Chemical Society, 1960, 82, 1132); Meticlane (France) M2790 and 1,365,504); metolazone (which may be prepared as disclosed in U.S. Patent No. 3,360,518); Paraflutizide (prepared as disclosed in Belgian Patent No. 620,829) polythiazide (which may be prepared as disclosed in U.S. Patent No. 3,009,911); Quinetazone (which may be prepared as disclosed in U.S. Patent No. 2,976,289); teclothiazide (which may be prepared as disclosed in U.S. Patent No. 2,976,289); Close et al., Journal of the American Chemical Society, 1960, 82, 1132) and trichlormethiazide (deStevens et al., Experientia, 1960, 16, 113); may be prepared as disclosed in) The disclosures of all such US patents are incorporated herein by reference.

Diuretic sulfonamide derivatives within the scope of the present invention include, but are not limited to: acetazolamide (which may be prepared as disclosed in US Pat. No. 2,980,679); ambuside (which may be prepared as disclosed in U.S. Patent No. 3,188,329); azocemide (which may be prepared as disclosed in U.S. Patent No. 3,665,002); bumetanide (which may be prepared as disclosed in U.S. Patent No. 3,634,583); butazolamide (which may be prepared as disclosed in British Patent No. 769,757); chloraminophenamide (which may be prepared as disclosed in U.S. Patent Nos. 2,809,194, 2,965,655 and 2,965,656); clofenamide (which may be prepared as disclosed in Olivier, Rec. Trav. Chim., 1918, 37, 307); clofamide (which may be prepared as disclosed in U.S. Patent No. 3,459,756); chlorexolone (which may be prepared as disclosed in U.S. Patent No. 3,183,243); disulfamide (which may be prepared as disclosed in British Patent No. 851,287); ethoxolamide (which may be prepared as disclosed in British Patent No. 795,174); furosemide (which may be prepared as disclosed in U.S. Patent No. 3,058,882); mepruside (which may be prepared as disclosed in U.S. Patent No. 3,356,692); metazolamide (which may be prepared as disclosed in U.S. Patent No. 2,783,241); pyrethanide (which may be prepared as disclosed in U.S. Patent No. 4,010,273); toracemide (which may be prepared as disclosed in U.S. Patent No. 4,018,929); tripamide (which may be prepared as disclosed in Japanese Patent No. 73 05,585); and Zipamide (which may be prepared as disclosed in U.S. Patent No. 3,567,777). The disclosures of all such US patents are incorporated herein by reference.

Other additional agents for combination with the compounds disclosed herein include PCSK9 translation inhibitors described in WO2016/055901, which is incorporated herein by reference.

In some embodiments, the additional therapeutic agent is an HMG-CoA reductase inhibitor, an HMG-CoA synthetase inhibitor, an HMG-CoA reductase gene expression inhibitor, an HMG-CoA synthetase gene expression inhibitor, an MTP/apo B secretion inhibitor, a CETP inhibitor, Bile acid absorption inhibitor, cholesterol absorption inhibitor, cholesterol synthesis inhibitor, squalene synthase inhibitor, squalene epoxidase inhibitor, squalene cyclase inhibitor, squalene epoxidase/squalene cyclase combination inhibitor, fibrate, niacin, combination of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid sequestrants and PCSK9 translation inhibitors.

For the treatment of sepsis or septic shock, the disclosed methods may include additional therapeutic agents such as antibiotics. In certain embodiments, the antibiotic is an aminoglycoside such as, but not limited to, kanamycin, amikacin, tobramycin, dibecacin, gentamicin, sisomycin, netylmycin, streptomycin, and neomycin B , C and E. Other antibiotics include carbapenems such as, but not limited to, imiphenum, meropenum, ertapenem, dorifenum, panifenum, viapenem, rasupenum, tebifenum, lenafenum, tomophenum, and tienphenum. includes

Additional antibiotics include quinoxolones such as, but not limited to, ciprofloxacin, garenoxacin, gatifloxacin, gemifloxacin, levofloxacin, synoxacin, nalidixic acid, moxifloxacin, oxolinic acid, pyromidic acid , pipemidic acid, rosoxacin, enoxacin, floxacin, romefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, lufloxacin, valofloxacin, grepafloxacin, levofloxacin, Pajufloxacin, Spafloxacin, Temfloxacin, Tosufloxacin, Clinafloxacin, Cytafloxacin, Trovafloxacin, Prulifloxacin, Derafloxacin, JNJ-Q2, Nemonoxacin, and Zavoflox includes the reaper

Additional antibiotics include tetracyclines such as, but not limited to, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, metacycline, minocycline, lolitetracycline and tigecycline.

Additional antibiotics useful in the methods disclosed herein include, but are not limited to, ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol, and ticarcillin.

pharmaceutical composition

The compositions and methods of the present invention can be used to treat a subject in need thereof. In certain embodiments, the subject is a mammal, such as a human, or a non-human mammal. When administered to a subject, such as a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. do. In a preferred embodiment, when such pharmaceutical compositions are intended for human administration, in particular for invasive routes of administration (i.e. routes that avoid transport or diffusion through the epithelial barrier, such as injection or implantation), the aqueous solution is free or substantially free of pyrogens. with no pyrogenic substances. The excipient may be selected, for example, to delay release of the agent or to selectively target one or more cells, tissues or organs. Pharmaceutical compositions may be in dosage unit form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilisates for reconstitution, powders, solutions, syrups, suppositories, injections, and the like. The composition may also be presented as a transdermal delivery system, eg, a skin patch. The compositions may also be in solutions suitable for topical administration, such as eye drops.

A pharmaceutically acceptable carrier may contain, for example, a physiologically acceptable agent that acts to stabilize a compound, such as a compound of the invention, increase solubility of the compound, or increase absorption of the compound. . Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizing agents or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The manufacture of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix, into which, for example, a compound of the invention is incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable and metabolizable carriers that are relatively simple to prepare and administer.

The phrase "pharmaceutically acceptable" herein is, within the scope of sound medical judgment, suitable for use in contact with the tissue of a subject without undue toxicity, irritation, allergic reaction, or other problems or complications, and has a reasonable benefit/risk ratio. risk ratio) to refer to a compound, substance, composition, and/or dosage form.

As used herein, the phrase “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the subject. Some examples of substances that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

Pharmaceutical compositions (formulations) can be prepared, for example, orally (e.g. as aqueous or non-aqueous solutions or suspensions in drenches, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (eg, sublingual); anus, rectum, or vagina (eg, a pessary, cream or foam); parenteral (including, for example, intramuscularly, intravenously, subcutaneously or intrathecally as a sterile solution or suspension); nasal cavity; intraperitoneal; subcutaneously; transdermal (eg, patches applied to the skin); and topical (eg, creams, ointments or sprays, or eye drops applied to the skin). The compounds may also be formulated for inhalation. In certain embodiments, the compound may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in US Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, and patents cited therein. can be found in

The formulations may conveniently be presented in unit dosage form and may be prepared by any method well known in the pharmaceutical art. The amount of active ingredient that may be combined with the carrier materials to prepare a single dosage form will vary depending upon the subject being treated and the particular mode of administration. The amount of active ingredient that may be combined with the carrier materials to prepare a single dosage form will generally be that amount of the composition to produce a therapeutic effect. Generally, out of 100%, this amount will range from about 1% to about 99%, preferably from about 5% to about 70%, and most preferably from about 10% to about 30% of the active ingredient.

Methods of preparing these formulations or compositions comprise the step of bringing into association an active compound, such as a compound of the invention, with a carrier and optionally one or more accessory ingredients. In general, formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with a liquid carrier, or finely divided solid carrier, or both, and then, if necessary, shaping the product.

Formulations of the present invention suitable for oral administration include capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (flavored bases, usually using sucrose and acacia or tragacanth). , in the form of lyophilisates, powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, with pastilles (inert bases such as gelatin and glycerin) , or using sucrose and acacia) and/or mouthwash, each containing a predetermined amount of a compound of the present invention as an active ingredient. The composition or compound may also be administered as a bolus, electuary or paste.

For the preparation of solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient may be formulated in one or more pharmaceutically acceptable carriers. , eg, sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders such as carboxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) moisturizing agents such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retardants such as paraffin; (6) absorption enhancers such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents such as modified and unmodified cyclodextrins; and (11) colorants. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also include a buffer. Solid compositions of a similar type may be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

Tablets may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets use binders (e.g., gelatin or hydroxypropylmethyl cellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfactants or dispersants can be manufactured. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

Tablets, and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, are optionally used in coatings and shells such as enteric coatings and pharmaceutical formulations. It can be scored or prepared with other well-known coatings. They may also be formulated to provide slow or controlled release of the internal active ingredient using, for example, hydroxypropylmethyl cellulose in various proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. can They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating the sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water, or some other sterile injectable medium immediately prior to use. These compositions may also optionally contain opacifying agents and may be of a composition which releases the active ingredient(s) alone, or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate together with one or more of the aforementioned excipients.

Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophilisates for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, Ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol , fatty acid esters of polyethylene glycol and sorbitan, and mixtures thereof.

In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

Suspensions may contain, in addition to the active compound, suspending agents, for example ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and their may contain mixtures.

Formulations of pharmaceutical compositions for rectal, vaginal, or urethral administration may be presented as suppositories, which contain one or more active compounds in one or more suitable non-irritating excipients or carriers (e.g., cocoa butter, polyethylene glycol, suppository waxes). or salicylates), which are solid at room temperature but liquid at body temperature, and will dissolve in the rectum or vaginal cavity to release the active compound.

Formulations of pharmaceutical compositions for oral administration may be presented as mouthwashes, or oral sprays, or oral ointments.

Alternatively or additionally, the composition may be formulated for delivery through a catheter, stent, wire, or other intraluminal device. Delivery via such a device may be particularly useful for delivery to the bladder, urethra, ureter, rectum, or intestine.

Formulations suitable for vaginal administration also include pessary, tampon, cream, gel, paste, foam or spray formulations containing carriers as are known to be suitable in the art.

Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be admixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants as may be required.

Ointments, pastes, creams and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays may contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays may additionally contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

Transdermal patches have the added advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in an appropriate medium. Absorption enhancers may also be used to increase the flux of the compound through the skin. The rate of this flow can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions, and the like are also contemplated as being within the scope of this invention. Exemplary ophthalmic formulations are described in US Publication Nos. 2005/0080056, 2005/0059744, 2005/0031697 and 2005/004074 and US Pat. No. 6,583,124, the contents of which are incorporated herein by reference. do. If desired, the liquid ophthalmic formulation has properties similar to, or is compatible with, tears, aqueous humor, or vitreous humor. A preferred route of administration is topical administration (eg, topical administration such as eye drops, or administration via an implant).

As used herein, the phrases “parenterally administered” and “parenterally administered” refer to modes of administration other than enteral and topical administration, usually by injection, and include intravenous, intramuscular, intraarterial, intrathecal intra, intracystic, intraorbital, intracardiac, intradermal, intraperitoneal, bronchial, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intravertebral, and intrasternal injections and infusions. it is not Pharmaceutical compositions suitable for parenteral administration may be reconstituted into one or more pharmaceutically acceptable isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions of one or more active compounds, or sterile injectable solutions or dispersions immediately prior to use. It may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (eg, glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

In addition, these compositions may contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like in the composition. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption, such as aluminum monostearate and gelatin.

In some instances, in order to prolong the effect of the drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of such a drug then depends on its rate of dissolution, which in turn may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is achieved by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms are prepared by forming microcapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by entrapping these drugs in liposomes or microemulsions that are compatible with body tissues.

For use in the methods of the present invention, the active compound by itself or, for example, in combination with a pharmaceutically acceptable carrier, is a pharmaceutical containing 0.1 to 99.5% (more preferably 0.5 to 90%) of active ingredient. It may be provided as a composition.

The method of introduction may also be provided by a rechargeable or biodegradable device. Various sustained-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including proteinaceous biopharmaceuticals, in recent years. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants for sustained release of compounds at specific target sites.

Actual dosage levels of the active ingredient in a pharmaceutical composition may vary to obtain an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without toxicity to the patient.

The selected dosage level will depend on the activity of the particular compound or combination of compounds employed, or ester, salt or amide thereof, route of administration, time of administration, rate of excretion of the particular compound(s) employed, duration of treatment, and particular compound(s) employed. ) will depend on a variety of factors, including other drugs, compounds and/or substances used in combination with .

A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the required pharmaceutical composition. For example, a physician or veterinarian may begin a dose of the pharmaceutical composition or compound at a lower level than necessary to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant a concentration of a compound sufficient to produce the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary with the weight, sex, age, and medical history of the subject. Other factors affecting the effective amount may include, but are not limited to, the condition of the subject being treated, the severity of the disorder, the stability of the compound, and another type of therapeutic agent administered, if necessary, with the compound of the present invention. A larger total dose may be delivered by multiple administrations of an agent. Methods for determining efficacy and dosage are known in the art (Isselbacher et al . (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

In general, a suitable daily dose of active compound for use in the compositions and methods of the present invention will be that amount of compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

If desired, an effective daily dose of the active compound will be 1, 2, 3, 4, 5, 6 or more sub-administered individually, optionally in unit dosage form, at appropriate intervals over the course of the day. It can be administered in a dose. In certain embodiments of the invention, the active compound may be administered twice or three times a day. In a preferred embodiment, the active compound will be administered once daily.

In certain embodiments, the compounds of the present invention may be used alone or co-administered with another type of therapeutic agent. As used herein, the phrase “conjoint administration” refers to any dosage form of two or more different therapeutic compounds such that a second compound is administered while the previously administered therapeutic compound is still effective in the body (eg, Both compounds are effective simultaneously in a subject, which may include a synergistic effect of both compounds). For example, different therapeutic compounds may be administered concomitantly or sequentially in the same formulation or in separate formulations. In certain embodiments, the different therapeutic compounds may be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or 1 week of each other. Thus, subjects receiving such treatment may benefit from the combined effects of different therapeutic compounds.

In certain embodiments, co-administration of a compound of the present invention with one or more additional therapeutic agent(s) is each of the compound of the present invention (eg, a compound of Formula I or Ia) or one or more additional therapeutic agent(s) provides improved efficacy compared to individual administration of In certain embodiments thereof, co-administration provides an additive effect, wherein the additive effect refers to the sum of the respective effects of separate administration of a compound of the invention and one or more additional therapeutic agent(s).

The present invention encompasses the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the present invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, contemplated salts of the present invention include L-arginine, benentamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino) Ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1- (2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, contemplated salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts.

Pharmaceutically acceptable acid addition salts may also exist in various solvates, for example, solvates with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of these solvates may also be prepared. The source of such a solvate may be obtained from the solvent of crystallization, intrinsic to the preparation or solvent of crystallization, or may be added externally to such solvent.

In addition, wetting agents, emulsifying agents and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, mold release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) fat-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Example

Examples of compounds of formula (I) or pharmaceutically acceptable salts thereof having useful biological activity are listed in Tables 1-10. ^{The 1} H NMR spectrum is a self-shielded z-gradient coil for reverse phase detection, operating at 400 MHz (proton frequency), 5 mm 1H/nX wideband probehead, with transmitter offset frequency shift. It was performed on a Varian MR-400 spectrometer with a deuterium digital lock channel unit, a quadrature digital detection unit. Chemical shifts are reported in ppm as the δ value relative to trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in Hertz (Hz), and multiplicity is reported using the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, nd = undetermined).

A. Analysis method

Method 1 (Acid FA)

UPLC Settings

Solvent: - A Water containing 0.1% formic acid (high purity through PureLab Option unit)

B Acetonitrile with 0.1% (V/V) formic acid (far UV grade)

column: - Acquity UPLC HSS C18 1.8 um 100 x 2.1 mm. (Plus Guard Cartridge)

Flow rate: - 0.5 mL/min

gradient:

Injection 0.5-2 uL

UV Detection with Waters DAD

Starting range (nm) 210 Termination range (nm) 400 Resolution (nm) 1.2

MS detection: Waters SQD2, single quadrupole UPLC-MS

Scan range for MS data (m/z)

Start (m/z) 100

End (m/z) 700 or 1500 if needed

Use +ve / -ve toggle

Ionization is ESI.

ESI voltage and temperature are as follows:

Source 150C 3.5KV capillary 25V cone

Method 2 (Base FA)

UPLC Settings

Solvent: - Acetonitrile (Range UV Grade)

Water containing 10 mM ammonium bicarbonate (ammonium bicarbonate) (high purity through PureLab Option Unit)

column: - Acquity UPLC BEH Shield RP18 1.7 um 100 x 2.1 mm. (Plus Guard Cartridge)

Flow rate: - 0.5 mL/min

Gradient: - A: water / base B: MeCN / base

A typical infusion of 0.5-2 uL (concentration approx. 0.2-1 mg/mL).

UV Detection with Waters DAD

Starting range (nm) 210 Termination range (nm) 400 Resolution (nm) 1.2

Other wavelength traces are extracted from the DAD data.

MS detection: Waters SQD2, single quadrupole UPLC-MS

The flow splitter shows about 300 ul/min for the mass spectrum.

Scan range for MS data (m/z)

Start (m/z) 100

End (m/z) 700 or 1500 if needed

Use +ve / -ve toggle

Preparative Reversed Phase HPLC Conditions

Preparative HPLC

Waters Micromass ZQ/Sample manager 2767

photodiode array detector 2996;

Column: XTerra Prep MS C18 Column (5 μm, 19 x 150 mm, Waters)

Flow rate: 20 mL/min using MS detection

UV wavelength: 254 nm.

mobile phase: solvent A (water:MeCN:HCO2H 95:5:0.05); Solvent B (water:MeCN:HCO2H 5:95:0.05)

gradient:

Flash chromatography is performed using an Isolera MPLC system (manufactured by Biotage) using pre-filled silica gel or reversed-phase cartridges (provided by Biotage or Interchim).

B. Chemical Synthesis

The general procedures used in the methods for preparing the compounds of the present invention are described below:

Scheme 1

2-chloro-5- (methylthio) pyrimidine (100B)

Dimethyl disulfide (13.96 mL, 155.4 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (700 mL) at -75 °C with cooled 5-bromo-2-chloropyrimidine (100A) (30 g, 155.4 mmol, 1.0 eq). ) was added under nitrogen. To this mixture was added dropwise a solution of n -butyl lithium (2.5 M, 68.4 mL, 170.9 mmol, 1.0 eq) over 1.5 hours, maintaining the internal temperature between -70°C and -75°C throughout the addition. After the addition was complete, the mixture was stirred at -75° C. for 4.5 h, then quenched by slow addition of saturated ammonium chloride solution (100 mL). The cooling bath was removed and the reaction allowed to warm to room temperature under nitrogen over 18 hours. The reaction mixture was diluted with ethyl acetate (500 mL) and washed with water (50 mL) and then saturated brine (50 mL). The organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude pale yellow oil obtained was purified by flash chromatography (eluting with 0-50% ethyl acetate in isohexane) to afford the desired product 2-chloro-5-(methylthio)pyrimidine as a waxy light yellow solid (100B). did.

Yield: 7.39 g (29%). ¹ H NMR (CDCl ₃ ) d 8.48 (2H, s), 2.54 (3H, s); MS (ESI+) m / z 161 (M+H) ⁺ .

General Method 1

tert -Butyl (2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C)

tert -Butyl(3-amino-2-methylpropyl)carbamate (4.79 g, 25.52 mmol, 1.05 eq) was dissolved in 2-chloro-5-(methylthio)pyrimidine (100B) in anhydrous dimethylformamide (50 mL). (3.89 g, 24.31 mmol, 1.0 eq) and cesium carbonate (11.85 g, 36.46 mmol, 1.4 eq) were added to a stirred suspension. The mixture was stirred for 18 h at room temperature and then concentrated under reduced pressure to about 20 mL. The liquid was diluted with ethyl acetate (100 mL), washed with water (75 mL) and brine (50 mL), then dried over magnesium sulfate. The solvent was removed in vacuo to give a crude residue, which was purified by flash chromatography (eluting with 0-50% ethyl acetate in isohexane) to give the desired product tert -butyl(2-methyl-3-((5-( Methylthio)pyrimidin-2-yl)amino)propyl) carbamate (100C) was obtained as a pale yellow oil.

Yield: 6.59 g (86%). MS (ESI+) m / z 313 (M+H) ⁺ .

tert -Butyl (S)-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl) carbamate (100D)

Racemic tert -butyl(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C) (5 g) was prepared by chiral SFC using the following conditions Purified: YMC amylose-C 30/70 MeOH/CO ₂ , 100 mL/min, 120 bar, 40° C., GLS 40 psi, system 3900 psi, drop 140 bar, Stacker, DAD 245 nm.

First eluting isomer, 1.3 min. tert -Butyl ( R )-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate

Yield: 2.05 g. ¹H NMR (400 MHz, CDCl ₃ ) d 8.34 (s, 2H), 5.79 (dd, J =6.7, 6.7 Hz, 1H), 5.18 (dd, J =5.8, 5.8 Hz, 1H), 3.50 - 3.39 (m) , 1H), 3.33 - 3.14 (m, 2H), 3.04 - 2.94 (m, 1H), 2.36 (s, 3H), 1.94 - 1.85 (m, 1H), 1.45 (s, 9H), 0.95 (d, J) =6.9 Hz, 3H). MS (ESI+) m / z 313 (M+H) ⁺ .

Second eluting isomer, 1.7 min. tert -Butyl ( S )-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate ( 100D )

Yield: 2.48 g. ¹H NMR (400 MHz, CDCl ₃ ) d 8.34 (s, 2H), 5.77 - 5.69 (m, 1H), 5.15 (dd, J = 6.5, 6.5 Hz, 1H), 3.50 - 3.40 (m, 1H), 3.33 - 3.16 (m, 2H), 3.03 - 2.94 (m, 1H), 2.36 (s, 3H), 1.95 - 1.85 (m, 1H), 1.47 (s, 9H), 0.95 (d, J = 6.9 Hz, 3H) ); MS (ESI+) m / z 313 (M+H) ⁺ .

general method 2

( R )-2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E)

A solution of hydrogen chloride (15 mL, 4 M in 1,4-dioxane) was mixed with tert -butyl ( S )-(2-ethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl Carbamate (100D) (600 mg, 2.48 mmol) was added, and the mixture was stirred at room temperature for 1 h The solvent was removed in vacuo to obtain the desired product ( R )-2-methyl- N ¹ -(5-(methyl Thio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) was obtained as a pale yellow semi-solid.

Yield: 548 mg (100%) HCl salt. ¹H NMR (400 MHz, DMSO) d 8.39 (s, 2H), 8.00 - 7.96 (m, 3H), 7.72 - 7.72 (m, 1H), 3.32 - 3.18 (m, 2H), 2.88 - 2.80 (m, 1H) ), 2.66 - 2.55 (m, 1H), 2.38 (s, 3H), 2.13 - 2.00 (m, 1H), 0.96 (d, J =6.8 Hz, 3H); MS (ESI+) m / z 213 (M+H) ⁺ .

common method 3

ethyl ( S )-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylate (100F)

Ethyl-2-chlorobenzothiazole-6-carboxylate (600 mg, 2.48 mmol, 1.0 eq) in anhydrous dimethylformamide (20 mL) under nitrogen ( R )-2-methyl- N ¹ -(5- To a stirred solution of (methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (548 mg, 2.48 mmol, 1.0 eq) and triethylamine (1.73 mL, 12.44 mmol, 5.0 eq) added. The mixture was stirred at room temperature for 48 h and then concentrated in vacuo to about 5 mL. Water (25 mL) was added and the mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL), then dried over magnesium sulfate. The solvent was removed in vacuo to give a crude yellow oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to the desired ethyl ( S )-2-((2-methyl-3-( (5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylate (100F) was obtained as an off-white solid.

Yield: 647 mg (62%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.37 (s, 2H), 8.27 (d, J =1.5 Hz, 1H), 7.99 (dd, J =1.8, 8.5 Hz, 1H), 7.51 (d, J =8.4) Hz, 1H), 7.02 - 6.98 (m, 1H), 5.83 (dd, J =6.7, 6.7 Hz, 1H), 4.38 (q, J =7.2 Hz, 2H), 3.67 - 3.47 (m, 2H), 3.42 - 3.24 (m, 2H), 2.37 (s, 3H), 2.20 - 2.11 (m, 1H), 1.40 (dd, J =7.2, 7.2 Hz, 3H), 1.07 (d, J =7.0 Hz, 3H); MS (ESI+) m / z 418 (M+H) ⁺ .

common method 4

( S )-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylic acid (Example 1)

Lithium hydroxide monohydrate (318 mg, 7.75 mmol, 5.0 eq) was mixed with ethyl ( S )-2-((2-methyl-3-((5-(methylthio)) in ethanol (7 mL) and water (5 mL). To a stirred solution of pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylate (100F) (647 mg, 1.55 mmol, 1.0 eq). The mixture was stirred for 18 h at ambient temperature and then concentrated under reduced pressure. Water (5 mL) was added to the residue and the mixture was acidified to pH about 3 with aqueous hydrochloric acid solution (2 M). The precipitate formed was collected by filtration, washed with water and dried under high vacuum to obtain the desired product ( S )-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino )propyl)amino)benzo[ d ]thiazole-6-carboxylic acid (Example 1) was obtained as a pale yellow solid.

common method 5

methyl ( S )-2-methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxamido)propanoate (100G)

1- [bis (dimethylamino) methylene] -1 H -1,2,3- triazolo [4,5- b] pyridinium 3-oxide hexafluorophosphate (HATU, 146 mg, 0.385 mmol , 1.5 eq) (S )-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole in dimethylformamide (5 mL) -6-carboxylic acid (1) (100 mg, 0.257 mmol, 1.0 eq), triethylamine (0.36 mL, 2.57 mmol, 10.0 eq) and methyl-2-amino-2-methylpropanoate hydrochloride (196 mg, 1.28) mmol, 4.9 eq) and the reaction mixture is stirred at room temperature for 18 h. The solvent was removed under reduced pressure, and the obtained crude residue was purified by flash chromatography (eluting with 0-75% ethyl acetate in isohexane) to obtain the desired methyl ( S )-2-methyl-2-(2-((2) -Methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxamido)propanoate (G) was obtained as an off-white solid. did.

Yield: 120 mg (95%).¹H NMR (400 MHz, MeOD) d 8.34 (s, 2H), 8.10 (d, J =1.8 Hz, 1H), 7.77 (dd, J =1.9, 8.5 Hz, 1H) , 7.47 (d, J =8.5 Hz, 1H), 3.74 (s, 3H), 3.45 (dd, J =6.3, 22.4 Hz, 4H), 2.34 (s, 3H), 2.29 - 2.16 (m, 1H), 1.59 (s, 6H), 1.08 (d, J =6.9 Hz, 3H) N H- substituting protons were not observed; MS (ESI+) m / z 489 (M+H) ⁺ .

( S )-2-methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ] Thiazole-6-carboxamido) propanoic acid (Example 2)

Method similar to general method 4

Lithium hydroxide monohydrate (50 mg, 1.22 mmol, 5.0 eq) was mixed with methyl ( S )-2-methyl-2-(2-((2-methyl-3-((2-methyl-3-() of (5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxamido)propanoate (100G) (120 mg, 0.245 mmol, 1.0 eq) It was added to the stirred solution and the mixture was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was diluted with water (3 mL) and acidified to pH about 3 with an aqueous solution of hydrochloric acid (2 M). The precipitate formed was collected by filtration, washed with water and dried under high vacuum to afford the title compound as an off-white solid.

The following examples were synthesized using the above procedure:

[Table 1]

Scheme 2

general method 6

2-chloro- N -(4-methoxybenzyl)benzo[ d ]thiazole-6-sulfonamide (101B).

(4-methoxyphenyl)methanamine (134 mg, 0.97 mmol, 1.05 eq) was dissolved in 2-chlorobenzothiazole-6-sulfonyl chloride (101A) (250 mg, 0.932 mmol, 1.0 eq), triethylamine (0.39 mL, 2.79 mmol, 3.0 eq) was added dropwise to an ice-cold solution, and the mixture was stirred at 0° C. for 1 hour. The reaction mixture was concentrated under reduced pressure to give a solid, which was washed with ice-cold water (10 mL), ice-cold tetrahydrofuran (10 mL) and then dried under vacuum to obtain the desired 2-chloro- N- (4-methoxybenzyl). )benzo[ d ]thiazole-6-sulfonamide was obtained as a white solid (101B).

Yield: 295 mg (85%). ¹H NMR (400 MHz, DMSO) d 8.56 (d, J = 1.6 Hz, 1H), 8.24 (s, 1H), 8.11 (d, J = 8.7 Hz, 1H), 7.90 (dd, J = 1.9, 8.7 Hz) , 1H), 7.10 (d, J = 8.8 Hz, 2H), 6.76 (d, J = 8.8 Hz, 2H), 3.98 (s, 2H), 3.67 (s, 3H); MS (ESI+) m / z 369 (M+H) ⁺ .

The intermediates in Table 2 were synthesized using conditions similar to those described for Intermediate 101B:

[Table 2]

( S )- N -(4-Methoxybenzyl)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl) amino)benzo[ d ]thiazole-6-sulfonamide (101C)

The methodology applied was similar to that described in General Method 3.

Yield: 298 mg, (69%). ¹H NMR (400 MHz, DMSO) d 8.56 (1H, d, J = 1.6 Hz), 8.24 (1H, s), 8.11 (1H, d, J = 8.7 Hz), 7.90 (1H, dd, J = 1.9, 8.7 Hz), 7.10 (2H, d, J = 8.8 Hz), 6.76 (2H, d, J = 8.8 Hz), 3.98 (2H, s), 3.67 (3H, s); MS (ESI+) m/z 545 (M+H) ⁺ .

( S )-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ] Thiazole-6-sulfonamide (Example 87)

Trifluoroacetic acid (5 mL) was dissolved in N- (4-methoxybenzyl)-2-((2-methyl-3-((5-(methylthio)pyrimidine-2- It was added dropwise to an ice-cooled solution of yl)amino)propyl)amino)benzo[ d ]thiazole-6-sulfonamide (101C) (250 mg, 0.459 mmol, 1.0 eq). The mixture was stirred on ice for 30 minutes and then allowed to warm to ambient temperature over 18 hours. The reaction mixture was diluted with dichloromethane (15 mL) and saturated sodium hydrogen carbonate (15 mL) was added slowly. The organic phase was separated, washed with brine (5 mL), dried over magnesium sulfate and then concentrated to dryness under reduced pressure. The obtained crude residue was purified by flash chromatography (eluting with 0-10% methanol in DCM) to obtain the desired ( S )-2-((2-methyl-3-((5-(methylthio)pyrimidine-2) -yl)amino)propyl)amino)benzo[ d ]thiazole-6-sulfonamide (Example 87) was obtained as a white solid.

Yield: 190 mg (97%). ¹H NMR (400 MHz, DMSO) d 8.40 (dd, J = 5.5, 5.5 Hz, 1H), 8.34 (s, 2H), 8.12 (d, J = 1.6 Hz, 1H), 7.66 (dd, J = 2.0, 8.4 Hz, 1H), 7.51 (dd, J = 6.0, 6.0 Hz, 1H), 7.44 (d, J = 8.5 Hz, 1H), 7.21 (s, 2H), 3.49 - 3.42 (m, 1H), 3.31 - 3.23 (m, 3H), 2.34 (s, 3H), 2.17 - 2.07 (m, 1H), 0.96 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 425 (M+H) ⁺ .

Using the procedure described in Scheme 2 , according to General Method 3 , the following examples were prepared:

[Table 3]

Scheme 3

2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A)

The applied methodology was similar to the general method 2.

A solution of hydrogen chloride (54 mL, 4 M in 1,4-dioxane) was mixed with tert -butyl (2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl carbamate (100C ) (4.25 g, 13.60 mmol), and the mixture is stirred at room temperature for 1 h The solvent is removed under vacuum to the desired 2-methyl- N ¹ -(5-(methylthio)pyrimidin-2-yl) Propane-1,3-diamine hydrochloride (102A) was obtained as a pale yellow semi-solid The semi-crude sample was used in the next reaction without further purification.

N ^One -(6-bromobenzo[ d ]thiazol-2-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B)

The applied methodology was similar to the general method 3.

6-Bromo-2-chlorobenzothiazole (828 mg, 3.33 mmol, 0.95 eq) was dissolved in 2-methyl- N ¹ -(5-(methylthio)pyrimidine- in anhydrous dimethylformamide (25 mL) under nitrogen 2-yl)propane-1,3-diamine hydrochloride (102A) (1.00 g, 3.51 mmol, 1.0 eq) and cesium carbonate (3.43 g, 10.52 mmol, 3.0 eq) were added to a stirred suspension. The mixture was stirred at room temperature for 72 h and then concentrated in vacuo to about 3 mL. Water (25 mL) was added and the mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL), then dried over magnesium sulfate. The solvent was removed in vacuo to give a crude yellow oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) for the desired N ¹ -(6-bromobenzo[ d ]thiazole-2). -yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B) was obtained as a sticky yellow solid.

Yield: 266 mg (18%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.37 (s, 2H), 7.66 (d, J = 0.6 Hz, 1H), 7.37 (d, J = 1.9 Hz, 2H), 6.58 (s, 1H), 5.69 ( dd, J = 6.3, 6.3 Hz, 1H), 3.67 - 3.23 (m, 4H), 2.37 (s, 3H), 2.17 - 2.09 (m, 1H), 1.06 (d, J = 6.9 Hz, 3H).

General Method 7

tert -Butyl 4-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazol-6-yl)-1 H -Pyrazole-1-carboxylate (102C)

N ¹ -(6-bromobenzo[ d ]thiazol-2-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B) ) (50 mg, 0.12 mmol, 1.0 eq) with water (0.20 mL) and N, N a solution under nitrogen of a-dimethylformamide (0.80 mL) of (1- (tert-butoxycarbonyl) -1 H-pyrazol Zol-4-yl)boronic acid (27.8 mg, 0.13 mmol, 1.0 eq), cesium carbonate (58 mg, 0.18 mmol, 1.5 eq) and tetrakis(triphenylphosphine)palladium(0)(7 mg, 0.01 mmol) , 0.05 eq) was added to the solution. The reaction mixture was heated to 90° C. for 16 h. (1- (tert - butoxycarbonyl) -1 H - pyrazol-4-yl) boronic acid (27.8 mg, 0.13 mmol, 1.0 eq) and tetrakis (triphenylphosphine) palladium (0) (7 mg , 0.01 mmol, 0.05 eq) were added to the reaction and the mixture was heated to 90° C. under nitrogen for a further 16 h. The solvent was removed under reduced pressure; Water (2 mL) was added and the mixture was extracted with ethyl acetate (3×5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), dried through a phase separator and concentrated in vacuo. The obtained crude residue was purified by flash chromatography (eluting with 0-75% ethyl acetate in isohexane) to the desired tert- butyl 4-(2-((2-methyl-3-((5-(methylthio) pyrimidin-2-yl) amino) propyl) amino) benzo [d] thiazol-6-yl) -1 H - pyrazole-1-carboxylate (102C) was obtained as a solid mibaeksaek. The semi-crude product was used in the next reaction without further purification.

N ^One -(6-(1) H -Pyrazol-4-yl)benzo[ d ]thiazol-2-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 97)

A solution of hydrogen chloride (2 mL, 4 M in 1,4-dioxane) was mixed with tert -butyl 4-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino ) propyl) amino) benzo [d] thiazol-6-yl) -1 h - pyrazole-1-carboxylate (102C) (100 mg, 0.12 mmol) was added and stirred for 1 hour at room temperature in a mixture did. The solvent is removed in vacuo to remove the desired N ¹ -(6-(1 H -pyrazol-4-yl)benzo[ d ]thiazol-2-yl)-2-methyl- N ³ -(5-(methylthio) Pyrimidin-2-yl)propane-1,3-diamine (Example 97) was obtained as an off-white solid.

Using the procedure described in Scheme 3 , according to General Method 7 , the following examples were prepared:

[Table 4]

general method 8

2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)- N ³ -(6-(oxazol-2-yl)benzo[ d ]thiazol-2-yl)propane-1,3-diamine (Example 101)

N ¹ -(6-bromobenzo[ d ]thiazol-2-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B) ) (90 mg, 0.21 mmol, 1.0 eq) in N , N -dimethylformamide (0.80 mL) under nitrogen (0.044 mL, 0.21 mmol, 1.0 eq) and 2-(tri-n -butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 eq) and To a solution of tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol, 0.1 eq). The reaction mixture was heated to 90° C. for 16 h. An additional aliquot of 2-(tri- n -butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 eq) and tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol, 0.1 eq) was added to the reaction mixture and heated to 110° C. for 18 h. The reaction was cooled to room temperature, diluted with water (2 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), filtered through a pad of celite, dried through a phase separator and concentrated to dryness in vacuo. The obtained crude residue was purified twice by flash chromatography (first eluting with 0-100% ethyl acetate in isohexane, then ethyl acetate with 0-10% methanol) to give a semi-crude residue, This was further purified by reverse phase preparative HPLC to the desired 2-methyl- N ¹ -(5-(methylthio)pyrimidin-2-yl) -N ³ -(6-(oxazol-2-yl)benzo[ d ] Thiazol-2-yl)propane-1,3-diamine (101) was obtained as an off-white solid.

Using the procedure described in Scheme 3 , according to General Method 8 , the following examples were prepared:

[Table 5]

Scheme 4

Using the procedure described in Scheme 4 , according to General Method 3 , the following examples were prepared:

[Table 6]

Scheme 7

tert- Butyl (2,2-dimethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (105A)

The methodology applied was similar to that described in General Method 1 , except that additional heating was required.

tert -Butyl (3-amino-2,2-dimethylpropyl)carbamate (0.13 g, 0.65 mmol, 1.05 eq) was dissolved in 2-chloro-5-(methylthio ) in anhydrous N , N-dimethylformamide (1.5 mL). ) was added to a stirred suspension of pyrimidine (100B) (0.10 g, 0.62 mmol, 1.0 eq) and cesium carbonate (0.24 g, 0.75 mmol, 1.2 eq) and the mixture was stirred at 80° C. for 4 h. The reaction mixture was concentrated in vacuo, diluted with ethyl acetate (20 mL), washed with water (7.5 mL) and brine (5.0 mL), then dried through a phase separator. The solvent was removed in vacuo to afford the desired tert- butyl (2,2-dimethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate ( 105A ) as a pale yellow oil.

Yield: 0.163 g (81%). ¹H NMR (400 MHz, DMSO) d 8.37 (s, 2H), 7.24 (dd, J = 6.6, 6.6 Hz, 1H), 6.93 (dd, J = 6.3, 6.3 Hz, 1H), 3.20 (d, J = 6.8 Hz, 2H), 2.85 (d, J = 8.7 Hz, 2H), 2.40 (s, 3H), 1.43 (d, J = 3.3 Hz, 9H), 0.83 (s, 6H).

2,2-dimethyl- N ^One -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (105B)

The methodology applied was similar to that described in General Method 2.

A solution of hydrogen chloride (5 mL, 4 M in 1,4-dioxane) was mixed with tert- butyl (2,2-dimethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carba Mate ( 105A ) (0.16 g, 0.50 mmol) was added and the mixture was stirred at room temperature for 1 h. The solvent was removed in vacuo to afford the desired 2,2-dimethyl-N ¹ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride ( 105B ) as a pale yellow semi-solid.

Yield: 0.13 g (100%) HCl salt. ¹H NMR (400 MHz, DMSO) d 8.39 (s, 2H), 7.95 (s, 3H), 7.70 (s, 1H), 3.25 (d, J = 5.5 Hz, 2H), 2.68 - 2.61 (m, 2H) , 2.38 (s, 3H), 0.96 (s, 6H).

N 1-(benzo[ d ]oxazol-2-yl)-2,2-dimethyl- N 3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 164)

The methodology applied was similar to that described in General Method 3 , except that cesium carbonate was used as the common base.

2-Chlorobenzoxazole (0.06 mL, 0.54 mmol, 0.1 eq) was dissolved in 2,2-dimethyl-N ¹ -(5-(methylthio)pyrimidine- in anhydrous N,N -dimethylformamide (2.0 mL) under nitrogen 2-yl)propane-1,3-diamine hydrochloride ( 105B ) (0.13 g, 0.49 mmol, 1.0 eq) and cesium carbonate (0.48 g, 1.48 mmol, 3.0 eq) was added to a stirred solution. The mixture was stirred at 80° C. or room temperature for 16 h and then concentrated in vacuo. Water (2.5 mL) was added and the mixture was extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), then dried through a phase separator. The solvent was removed in vacuo to give a crude yellow oil, which was purified by preparative HPLC for the desired N ¹ -(benzo[ d ]oxazol-2-yl)-2,2-dimethyl- N ³ -(5-( Methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 164) was obtained as an off-white solid.

Using the procedure described in Scheme 7 , according to General Method 3 , the following examples were prepared:

[Table 9]

Scheme 8

3-chloro- N -(2-chlorobenzo[ d ]thiazol-6-yl)propane-1-sulfonamide (106B)

Sodium hydride (dispersion 60% in mineral oil) (326 mg, 8.15 mmol, 3.0 eq) of N, N - 2- chloro-benzothiazol-6-amine in dimethylformamide (25 mL) (500 mg, 2.71 mmol, 1.0 eq) of an ice-cold solution was added in small portions, and the mixture was stirred under ice cooling for 1 h. A solution of 3-chloropropane-1-sulfonyl chloride (673 mg, 3.80 mmol, 1.4 eq) in N, N -dimethylformamide (3 mL) was added dropwise, then the reaction mixture was warmed to ambient temperature over 3 h. made to be The reaction mixture was diluted with brine (20 mL) and extracted with ethyl acetate (2×25 mL). The combined organic fractions were combined and concentrated under reduced pressure to give a pale yellow oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to the desired product 3-chloro- N- (2-chlorobenzo [ d ] Thiazol-6-yl) propane-1-sulfonamide 106B was obtained as an off-white gum.

Yield: 427 mg (48%). ¹H NMR (400 MHz, MeOD) d 7.87 - 7.84 (m, 2H), 7.38 (dd, J = 2.3, 8.8 Hz, 1H), 3.67 (dd, J = 6.3, 6.3 Hz, 2H), 2.27 - 2.19 ( m, 2H). NOTE CH ₂ protons are not noticed by MeOD.

2-(2-chlorobenzo[ d ]thiazol-6-yl)isothiazolidine 1,1-dioxide (106C)

Sodium hydride (60% dispersion in mineral oil) (98 mg, 2.46 mmol, 2.0 eq) was dissolved in 3-chloro-N- (2-chlorobenzo[ d ]thiazole- in N , N -dimethylformamide (5 mL)- 6-yl)propane-1-sulfonamide (106B) (400 mg, 1.23 mmol, 1.0 eq) was added to an ice cold solution. The reaction mixture was stirred under ice cooling for 1 h, then quenched by careful addition of saturated ammonium chloride solution (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 20 mL), and the combined organic phases were washed with water (20 mL), brine (20 mL) and then concentrated in vacuo to give a gum. The crude product was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to afford the desired product (106C) as an off-white gum.

Yield: 220 mg (62%). ¹H NMR (400 MHz, CDCl ₃ ) d 7.92 (d, J = 8.9 Hz, 1H), 7.72 (d, J = 2.1 Hz, 1H), 7.38 (dd, J = 2.4, 8.9 Hz, 1H), 3.84 ( dd, J = 6.5, 6.5 Hz, 2H), 3.43 (dd, J = 7.5, 7.5 Hz, 2H), 2.63 - 2.55 (m, 2H).

2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazol-6-yl)isothiazolidine 1,1-dioxide (Example 175)

The methodology applied was similar to that described in General Method 3.

Yield: 25 mg (15%). ¹H NMR (400 MHz, DMSO) d 8.34 (s, 2H), 8.03 (dd, J = 5.6, 5.6 Hz, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.51 (dd, J = 5.9, 5.9 Hz, 1H), 7.35 (d, J = 8.8 Hz, 1H), 7.14 (dd, J = 2.4, 8.7 Hz, 1H), 3.72 (dd, J = 6.5, 6.5 Hz, 2H), 3.47 (dd, J = 7.5, 7.5 Hz, 2H), 3.45 - 3.38 (m, 1H), 3.30 - 3.22 (m, 3H), 2.44 - 2.36 (m, 2H), 2.35 (s, 3H), 2.13 - 2.06 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 465 (M+H) ⁺ .

Scheme 9

N -(2-chlorobenzo[ d ]thiazol-6-yl)methanesulfonamide (107B)

Methanesulfonyl chloride (0.055 mL, 0.706 mmol, 1.3 eq) was mixed with 2-chlorobenzothiazol-6-amine (100 mg, 0.54 mmol, 1.0 eq) and pyridine (0.066 mL, 0.815) in anhydrous dichloromethane (5 mL). mmol, 1.5 eq) was added dropwise to an ice-cold solution. The mixture was stirred at 0° C. for 15 min and then allowed to warm to ambient temperature over 1 h. The reaction mixture was quenched with water (1 mL). The organic phase was removed and concentrated under reduced pressure to give a pale yellow oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) for the desired N- (2-chlorobenzo[ d ]thiazole-6- 1) Methanesulfonamide (107B) was obtained as a pale yellow gum.

Yield: 135 mg (94.8%) ¹H NMR (400 MHz, CDCl ₃ ) d 7.88 (d, J = 8.8 Hz, 1H), 7.79 (d, J = 2.1 Hz, 1H), 3.00 (s, 3H). Aromatic H protons are _{not conspicuous by CDCl 3} , and no N H- substituting protons are observed.

N -(2-chlorobenzo[ d ]thiazol-6-yl)- N -methylmethanesulfonamide (107C)

Sodium hydride (60% dispersion in mineral oil) (31 mg, 0.772 mmol, 1.5 eq) was dissolved in N- (2-chlorobenzo[ d ]thiazol-6-yl)methanesulfonamide in anhydrous tetrahydrofuran (2 mL). (107B) (135 mg, 0.515 mmol, 1.0 eq) was added in small portions to an ice-cold solution. The mixture was stirred at room temperature for 2 hours. Iodomethane (0.048 mL, 0.772 mmol, 1.5 eq) was added and the mixture was stirred at room temperature for an additional 2 h. Water (1 mL) was added and the solvent was removed under high vacuum to give a pale yellow gum, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) N- (2-chlorobenzo[ d ]thiazole) -6-yl) -N -methylmethanesulfonamide (107C) was obtained as a pale yellow gum.

Yield: 100 mg (70%) ¹H NMR (400 MHz, CDCl ₃ ) d 7.95 (d, J = 8.8 Hz, 1H), 7.86 (d, J = 2.1 Hz, 1H), 7.48 (dd, J = 2.3, 8.8 Hz, 1H), 3.39 (s, 3H), 2.88 (s, 3H).

N -methyl- N -(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazol-6-yl)methanesulfonamide (Example 176)

The methodology applied was similar to that described in General Method 3.

Yield: 89 mg (60%) ¹H NMR (400 MHz, CDCl ₃ ) d 8.38 (s, 2H), 7.62 (d, J = 2.0 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.24 (d, J = 3.2 Hz, 1H), 5.74 (dd, J = 6.5, 6.5 Hz, 1H), 3.34 - 3.33 (m, 6H), 2.86 (s, 3H), 2.37 (s, 3H), 2.20 - 2.10 (m, 1H), 1.06 (d, J = 6.9 Hz, 3H). Not all NH- substituting protons were observed; MS (ESI+) m / z 453 (M+H) ⁺ .

Scheme 10

4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)pyrrolidin-2-one (108B)

The methodology applied was similar to that described in General Method 1.

5-Aminomethyl-pyrrolidin-2-one (108A) (1.0 g, 8.76 mmol, 1.0 eq) was mixed with 2-chloro-5- (methylthio) pyrimidine (100B) in anhydrous dimethylformamide (10 mL). (1.4 g, 8.76 mmol, 1.0 eq) and cesium carbonate (8.56 g, 26.28 mmol, 3.0 eq) were added to a stirred suspension. The mixture was heated to 50° C. for 18 h and then concentrated under reduced pressure. The obtained liquid was diluted with ethyl acetate (50 mL), washed with water (10 mL) and brine (10 mL), then dried through a phase separator. The solvent was removed under reduced pressure to give a crude residue, which was purified by trituration in methanol to obtain the desired product 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)pi Rollidin-2-one (108B) was obtained as a yellow solid. The aqueous phase is concentrated under reduced pressure, combined with the filtrate, and purified by flash chromatography (eluting with 0-10% methanol in dichloromethane) to the desired product 4-(((5-(methylthio)pyrimidin-2-yl) Amino)methyl)pyrrolidin-2-one (108B) was obtained as a yellow solid. The two crops were combined and used in the next step.

Yield: 0.74 g (36%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.35 (s, 2H), 5.79 - 5.76 (m, 1H), 5.48 (dd, J = 5.5, 5.5 Hz, 1H), 3.57 - 3.50 (m, 3H), 3.20 (dd, J = 5.3, 9.5 Hz, 1H), 2.89 - 2.84 (m, 1H), 2.53 - 2.46 (m, 1H), 2.37 (s, 3H), 2.16 (dd, J = 6.4, 17.1 Hz, 1H) ).

tert -Butyl 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C)

4-dimethylaminopyridine (5 mg, 0.04 mmol, 0.1 eq) was dissolved in dichloromethane (4.2 mL) with 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)pyrrolidine-2 A stirred suspension of -one (108B) (100 mg, 0.4 mmol, 1.0 eq), di- tert -butyl dicarbonate (229 mg, 1.0 mmol, 2.5 eq) and triethylamine (0.146 mL, 1.00 mmol, 2.5 eq) was added to The mixture was stirred at room temperature for 72 h, then water (3.0 mL) was added. The mixture was extracted with ethyl acetate (3 x 5.0 mL) and the combined organic phases washed with brine (2.5 mL), then dried through a phase separator. The solvent was removed under reduced pressure to give a crude residue, which was purified by reverse phase chromatography (0.1% formic acid solution in acetonitrile, eluting with 5-100%) to give the desired product tert -butyl 4-(((5-( Obtained methylthio))pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C).

Yield: 72 mg (53%). ¹H NMR (400 MHz, DMSO) d 8.35 (s, 2H), 7.62 (t, J = 6.1 Hz, 1H), 3.73 (dd, J = 7.8, 10.4 Hz, 1H), 3.50 - 3.44 (m, 1H) , 3.29 (d, J = 6.4 Hz, 2H), 2.60 - 2.55 (m, 2H), 2.36 (s, 3H), 2.28 (dd, J = 9.6, 20.9 Hz, 1H), 1.44 (s, 9H).

tert -Butyl (4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4-oxobutyl)carbamate (108D)

A solution of ammonium hydroxide (2.2 mL) was dissolved in tert -butyl 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C) (72 mg, 0.21 mmol) was added and the mixture was heated at 80° C. for 1.5 h. The mixture was cooled to room temperature and then extracted with dichloromethane (3 x 5 mL). The organic solvent was dried via a phase separator and then removed under reduced pressure to remove the desired product tert -butyl (4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4- Oxobutyl) carbamate (108D) was obtained.

Yield: 61 mg (86%). ¹H NMR (400 MHz, DMSO) d 8.33 (s, 2H), 7.32 (s, 1H), 7.25 (dd, J = 6.0, 6.0 Hz, 1H), 6.81 (s, 2H), 3.24 (dd, J = 6.2, 6.2 Hz, 2H), 2.97 (dd, J = 6.0, 6.0 Hz, 2H), 2.36 (s, 3H), 2.18 - 2.08 (m, 1H), 2.04 (d, J = 6.4 Hz, 2H), 1.38 (s, 9H).

4-amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride salt (108E)

The methodology applied was similar to that described in General Method 2.

Hydrogen chloride solution (0.7 mL, 4M in 1,4-dioxane) was mixed with tert -butyl (4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4-oxo Butyl)carbamate (108D) (61 mg, 0.17 mmol) was added and the mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure to afford the desired product 4-amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride salt (108E) as a pale yellow solid.

Yield: 56 mg (100%) HCl salt.

4-((6-( N -(4-methoxybenzyl)sulfamoyl)benzo[ d ]thiazol-2-yl)amino)-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (177)

The methodology applied was similar to that described in General Method 3.

2-Chloro- N- (4-methoxybenzyl)benzo[ d ]thiazole-6-sulfonamide (101B) (70 mg, 0.19 mmol, 1.10 eq) in anhydrous dimethylformamide (2.0 mL) under nitrogen 4 -amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride salt (108E) (56 mg, 0.17 mmol, 1.0 eq) and triethylamine (0.072 mL, 0.51 mmol, 3.0 eq) of a stirred solution. The mixture was stirred for 72 h at room temperature and then concentrated under reduced pressure. Water (2.5 mL) was added and the resulting precipitate was collected by filtration and then washed with methanol. The organic filtrate was concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography (eluting with 0-25% methanol in dichloromethane) to give the desired product 4-((6-( N- (4-methoxybenzyl) )sulfamoyl)benzo[ d ]thiazol-2-yl)amino)-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (Example 177) as an off-white solid was obtained as

Yield: 25 mg (25%). ¹H NMR (400 MHz, DMSO) d 8.45 (dd, J = 5.6, 5.6 Hz, 1H), 8.33 (s, 2H), 8.08 (d, J = 1.8 Hz, 1H), 7.88 (dd, J = 6.1, 6.1 Hz, 1H), 7.62 (dd, J = 1.9, 8.5 Hz, 1H), 7.49 - 7.42 (m, 2H), 7.39 - 7.37 (m, 1H), 7.14 (d, J = 8.7 Hz, 2H), 6.87 - 6.87 (m, 1H), 6.81 (d, J = 8.7 Hz, 2H), 3.89 (d, J = 6.0 Hz, 2H), 3.70 (s, 3H), 3.48 (d, J = 1.1 Hz, 2H) ), 3.37 (dd, J = 6.1, 6.1 Hz, 2H), 2.43 - 2.37 (m, 1H), 2.34 (s, 3H), 2.22 - 2.17 (m, 2H).

4-((5-(methylthio)pyrimidin-2-yl)amino)-3-(((6-sulfamoylbenzo[d]thiazol-2-yl)amino)methyl)butanamide (178)

4-((6-(N- (4-methoxybenzyl)sulfamoyl)benzo[ d ]thiazol-2-yl)amino)- in trifluoroacetic acid (0.3 mL) in anhydrous dichloromethane (0.3 mL) It was added dropwise to a cooled solution of 3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (177) (20 mg, 0.03 mmol, 1.0 eq) at 0°C. The mixture was stirred for 30 min and then allowed to warm to room temperature over 8 h. An additional aliquot of trifluoroacetic acid (1 mL) was added and the mixture was stirred for 16 h. The reaction mixture was concentrated under pressure and then carefully basified by addition of saturated aqueous sodium hydrogen carbonate solution (3 mL). The mixture was extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with brine (3 mL), dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The obtained crude residue was purified by flash chromatography (eluting with 0-20% methanol in dichloromethane) to obtain the desired product 4-((5-(methylthio)pyrimidin-2-yl)amino)-3-(( (6-sulfamoylbenzo[ d ]thiazol-2-yl)amino)methyl)butanamide (Example 178) was obtained as a white solid.

Yield: 12 mg (75%). ¹H NMR (400 MHz, DMSO) d 8.41 (dd, J = 5.5, 5.5 Hz, 1H), 8.33 (s, 2H), 8.13 (d, J = 1.8 Hz, 1H), 7.67 (dd, J = 1.9, 8.5 Hz, 1H), 7.47 - 7.43 (m, 2H), 7.38 (s, 1H), 7.21 (s, 2H), 6.86 (s, 1H), 3.46 - 3.46 (m, 2H), 3.36 (dd, J) = 6.2, 6.2 Hz, 2H), 2.39 - 2.38 (m, 1H), 2.34 (s, 3H), 2.21 - 2.17 (m, 2H).

The following examples were synthesized using the procedure described in Scheme 10:

[Table 10]

Scheme 11

4-((2-chlorobenzo[d]thiazol-6-yl)sulfonyl)morpholine (109B)

The methodology applied was similar to that described in General Method 6 using dichloromethane as solvent instead of tetrahydrofuran.

Yield: 686 mg. ¹H NMR (400 MHz, CDCl3) d 8.25 (d, J = 1.8 Hz, 1H), 8.10 (d, J = 8.6 Hz, 1H), 7.85 (dd, J = 1.3, 8.6 Hz, 1H), 3.75 (dd , J = 4.7, 4.7 Hz, 4H), 3.04 (dd, J = 4.7, 4.7 Hz, 4H); MS (ESI+) m/z 319 (M+H)+.

tert-Butyl (3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)-2-methylpropyl)carbamate (109D)

The methodology applied was similar to that described in General Method 1.

Yield: 433 mg. ¹H NMR (400 MHz, CDCl3) d 8.16 - 8.15 (m, 2H), 5.05 - 5.05 (m, 1H), 3.47 - 3.38 (m, 1H), 3.31 - 3.17 (m, 2H), 3.04 - 2.95 (m , 1H), 1.96 - 1.86 (m, 1H), 1.60 - 1.58 (m, 1H), 0.96 - 0.94 (m, 3H); MS (ESI+) m/z 333 (M+H)+.

N1-(5-(difluoromethoxy)pyrimidin-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E)

A solution of hydrogen chloride (2.7 mL, 4 M in 1,4-dioxane) was mixed with tert-butyl (3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)-2-methylpropyl)carba Mate (109D) (300 mg, 0.903 mmol) was added and stirred at room temperature for 15 min. The solvent was removed in vacuo to give the crude title compound N1-(5-(difluoromethoxy)pyrimidin-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E) for the next step without further purification did.

Yield: 225 mg (quantitative). MS (ESI+) m/z 233 (M+H)+.

N1-(5-(difluoromethoxy)pyrimidin-2-yl)-2-methyl-N3-(6-(morpholinosulfonyl)benzo[d]thiazol-2-yl)propane-1,3 -diamine (180)

The methodology applied was similar to that described in General Method 3.

Yield: 225 mg. ¹H NMR (400 MHz, CDCl3) d 8.21 (s, 2H), 7.97 (d, J = 1.5 Hz, 1H), 7.65 (dd, J = 1.8, 8.5 Hz, 1H), 7.59 (d, J = 8.5 Hz) , 1H), 6.91 (s, 1H), 6.43 (t, J = 71.6 Hz, 1H), 5.72 (dd, J = 6.6, 6.6 Hz, 1H), 3.77 - 3.73 (m, 4H), 3.62 - 3.51 ( m, 2H), 3.43 - 3.29 (m, 2H), 3.01 (dd, J = 4.6, 4.6 Hz, 4H), 2.21 - 2.12 (m, 1H), 1.08 (d, J = 6.9 Hz, 3H); (ESI+) m/z 515 (M+H)+.

Following the procedure described in Scheme 11 , the following examples were synthesized:

[Table 11]

Scheme 12

tert -Butyl (2-methyl-3- (pyridin-2-ylamino) propyl) carbamate (110B)

A solution of tert -butyl 3-amino-2-methylpropylcarbamate (110A) (150 mg, 0.80 mmol, 1.0 eq) in 1,4-dioxane (2 mL) was dissolved in 1,4-dioxane (10 mL), 2-bromopyridine (0.076 mL, 0.80 mmol, 1.0 eq), sodium tert -butoxide (383 mg, 3.98 mmol, 5.0 eq), 2-dicyclohexylphosphino-2′,4′,6 It was added to a solution of '-triisopropylbiphenyl (38 mg, 0.08 mmol, 0.1 eq) and tris(dibenzylideneacetone)dipalladium(0)(73 mg, 0.08 mmol, 0.1 eq). The reaction mixture was heated to 70° C. for 72 h. The solvent was removed under reduced pressure and the resulting residue was partitioned between water (2 mL) and ethyl acetate (5 mL). The mixture was filtered through celite, then the aqueous phase was removed and extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), dried through a phase separator, and then concentrated to dryness in vacuo. The obtained crude residue was purified by reverse phase chromatography (eluting with 5-95% acetonitrile in 10 mM aqueous ammonium bicarbonate solution) to obtain semi-purity of the desired tert -butyl (2-methyl-3-(pyridin-2-yl) Amino)propyl)carbamate (110B) was obtained as an off-white solid, which was used in the next step without further purification.

Yield: 27 mg (12%). MS (ESI+) m / z 266 (M+H) ⁺ .

2-methyl- N ^One -(pyridin-2-yl)propane-1,3-diamine hydrochloride (110C)

The methodology applied was similar to that described in General Method 2.

A solution of hydrogen chloride (0.4 mL, 4 M in 1,4-dioxane) was mixed with tert -butyl (2-methyl-3- (pyridin-2-ylamino) propyl) carbamate (108B) (27 mg, 0.10 mmol) , and the mixture was stirred at room temperature for 1 hour. The solvent was removed in vacuo to afford the desired product 2-methyl- N 1-(pyridin-2-yl)propane-1,3-diamine hydrochloride (110C) as a pale yellow semi-solid. The semi-crude sample was used in the next reaction without further purification.

Yield: 25 mg (% estimated quantitation).

2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)- N ³ -(pyridin-2-yl)propane-1,3-diamine (Example 187)

The methodology applied was similar to that described in General Method 3.

2-Chloro-5-methylsulfanyl-pyrimidine (100B) (17 mg, 0.11 mmol, 1.05 eq) was dissolved in 2-methyl-N ¹ -(pyridine ) in anhydrous N , N -dimethylformamide (0.5 mL) under nitrogen. -2-yl)propane-1,3-diamine hydrochloride (110C) (24 mg, 0.10 mmol, 1.0 eq) and cesium carbonate (99 mg, 0.30 mmol, 3.0 eq) was added to a stirred solution. The mixture was heated to 50° C. for 16 h and then concentrated in vacuo. Water (2 mL) was added and the mixture was extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), then dried through a phase separator. The solvent was removed in vacuo to give a crude yellow oil, which was purified by reverse phase chromatography (eluting with 5-95% acetonitrile in 10 mM aqueous ammonium bicarbonate solution) to give the desired 2-methyl- N ¹ -(5-(methyl Obtained thio)pyrimidin-2-yl) -N ³ -(pyridin-2-yl)propane-1,3-diamine (Example 187) as a sticky yellow solid.

Yield: 1.5 mg (5%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.35 (s, 2H), 8.10 (dd, J = 0.6, 3.6 Hz, 1H), 7.41 - 7.35 (m, 1H), 6.54 (dd, J = 5.3, 6.8 Hz) , 1H), 6.38 (d, J = 8.4 Hz, 1H), 5.93 - 5.93 (m, 1H), 4.96 - 4.96 (m, 1H), 3.53 - 3.21 (m, 4H), 2.35 (s, 3H), 2.10 - 2.01 (m, 1H), 1.03 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 290 (M+H) ⁺ .

Scheme 13

Methyl 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl) amino)pyrazine-2-carboxylate (188)

The methodology applied was similar to that described in General Method 3.

Methyl 5-bromopyrazine-2-carboxylate (114 mg, 0.53 mmol, 1.0 eq) was dissolved in 2-methyl-N ¹ -(5-(methyl ) in anhydrous N , N -dimethylformamide (2.0 mL) under nitrogen. Thio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A) (150 mg, 0.53 mmol, 1.0 eq) and cesium carbonate (514 mg, 1.58 mmol, 3.0 eq) was added to a stirred solution. The mixture was stirred at room temperature for 16 h and then concentrated in vacuo. Water (2.5 mL) was added and the mixture was extracted with ethyl acetate (3 x 5 mL). The combined organic phases were washed with water (2 mL) and brine (2.5 mL) and then dried through a phase separator. The solvent was removed in vacuo to give a brown oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane). The resulting semi-crude product was further purified by flash chromatography (eluting with 0-10% methanol in dichloromethanol) to the desired methyl 5-((2-methyl-3-((5-(methylthio)pyrimidine-2) -yl)amino)propyl)amino)pyrazine-2-carboxylate (Example 188) was obtained as a white solid.

Yield: 60 mg (32%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.76 (s, 1H), 8.37 (s, 2H), 7.91 (d, J = 1.4 Hz, 1H), 6.36 - 6.36 (m, 1H), 5.56 (dd, J = 6.0, 6.0 Hz, 1H), 3.95 (s, 3H), 3.60 - 3.51 (m, 2H), 3.40 - 3.26 (m, 2H), 2.38 (s, 3H), 2.10 - 2.03 (m, 1H), 1.04 (d, J = 6.9 Hz, 3H). MS (ESI+) m / z 349 (M+H) ⁺ .

5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (189)

The methodology applied was similar to that described in General Method 4.

Lithium hydroxide monohydrate (23 mg, 55 mmol, 5.0 eq) was mixed with methyl 5-((2-methyl-3-((5-(methylthio)pyrimidine-2) in ethanol (0.4 mL) and water (0.4 mL). -yl)amino)propyl)amino)pyrazine-2-carboxylate (188) (38 mg, 0.11 mmol, 1.0 eq) was added to a stirred solution. The mixture was stirred for 72 h at ambient temperature and then concentrated under reduced pressure. Water (0.5 mL) was added to the residue and the mixture was acidified to pH about 3 with aqueous hydrochloric acid (2 M). The sticky precipitate was collected by filtration, then extracted with ethyl acetate (3 x 3 mL), washed with water (1 mL) and dried through a phase separator. The solvent was removed in vacuo to give the desired 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (Example 189) to a pale yellow color. Obtained as a solid.

Yield: 34 mg (94%); ¹H NMR (400 MHz, DMSO) 8.52 (s, 1H), 8.36 (s, 2H), 7.99 (s, 2H), 7.65 - 7.64 (m, 1H), 3.39 - 3.20 (m, 4H), 2.36 ( s, 3H), 2.12 - 2.03 (m, 1H), 0.94 (d, J = 6.8 Hz, 3H), one N H proton not observed; MS (ESI+) m / z 335 (M+H) ⁺ .

(4-Hydroxypiperidin-1-yl)(5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl ) Methanone (Example 190)

The methodology applied was similar to that described in General Method 5.

1- [bis (dimethylamino) methylene] -1 H -1,2,3- triazolo [4,5- b] pyridinium 3-oxide hexafluorophosphate (HATU, 58 mg, 0.15 mmol , 1.5 eq) an N, N-dimethylformamide (1 mL) of 5 - ((2-methyl-3 - ((5- (methylthio) pyrimidin-2-yl) amino) propyl) amino) pyrazine-2-carboxylic acid ( 189) (34 mg, 0.10 mmol, 1.0 eq) and 4-hydroxypiperidine (103 mg, 1.02 mmol, 10 eq), and the reaction mixture was stirred at room temperature for 18 h. The solvent was removed under reduced pressure and the obtained crude residue was purified by reverse-phase preparative HPLC to the desired (4-hydroxypiperidin-1-yl)(5-((2-methyl-3-((5-( Methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl)methanone (Example 190) was obtained as an off-white solid.

Yield: 22 mg (53%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.46 (s, 1H), 8.37 (s, 2H), 7.79 (s, 1H), 5.97 - 5.97 (m, 1H), 5.71 - 5.71 (m, 1H), 4.15 - 4.15 (m, 2H), 4.01 - 3.94 (m, 1H), 3.58 - 3.47 (m, 2H), 3.42 - 3.25 (m, 4H), 2.37 (s, 3H), 2.08 (ddd, J = 11.5, 11.5, 5.1 Hz, 1H), 2.00 - 1.94 (m, 2H), 1.62 - 1.60 (m, 3H), 1.04 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 418 (M+H) ⁺ .

Scheme 16

5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-amine (111B)

The methodology applied was similar to that described in General Method 7.

2-Bromo-5-methyl-1,3,4-oxadiazole (0.58 g, 3.58 mmol, 1.05 eq) was dissolved in 5- in water (5.5 mL) and 1,4-dioxane (22.5 mL) under nitrogen (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (111A) (0.75 g, 3.41 mmol, 1.0 eq), cesium carbonate (3.33) g, 10.22 mmol, 3.0 eq) and tetrakis(triphenylphosphino)palladium(0) (0.39 g, 0.34 mmol, 0.10 eq). The reaction mixture was heated to 100° C. for 16 h. The solvent was removed in vacuo, water (20 mL) was added and the mixture was extracted with ethyl acetate (6×50 mL). The combined organic phases were washed with water (10 mL) and brine (10 mL), dried through a phase separator and concentrated in vacuo. The obtained crude residue was purified by reverse phase chromatography (eluting with 5-30% acetonitrile in 10 mM ammonium bicarbonate solution) to the desired 5-(5-methyl-1,3,4-oxadiazole-2- yl)pyridin-2-amine (111B) was obtained as an off-white solid.

Yield: 153 mg (25%). ¹H NMR (400 MHz, DMSO) d 8.49 (d, J = 2.0 Hz, 1H), 7.87 (dd, J = 2.4, 8.8 Hz, 1H), 6.75 (s, 2H), 6.57 (d, J = 8.8 Hz) , 1H), 2.53 (s, 3H).

tert -Butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)amino)propyl) carbamate (111C)

tert -Butyl N- (2-methyl-3-oxopropyl)carbamate (111B) (225 mg, 1.21 mmol, 1.2 eq) was dissolved in 5-(5-methyl-1,3, 4-oxadiazol-2-yl)pyridin-2-amine (114B) (153 mg, 1.00 mmol, 1.0 eq), acetic acid (0.230 mL, 4.02 mmol, 4.0 eq) and molecular sieve (type 4 Å, 250 mg) was added to the solution of The reaction mixture was stirred at room temperature for 5 min, then sodium triacetoxyborohydride (532 mg, 2.51 mmol, 2.5 eq) was added in one portion. The mixture was stirred at room temperature for 40 hours. tert -Butyl N- (2-methyl-3-oxopropyl)carbamate (225 mg, 1.21 mmol, 1.2 eq) was added and the reaction mixture was stirred at room temperature for a further 72 h. The reaction was quenched by careful addition of saturated aqueous sodium hydrogen carbonate solution (30 mL). The mixture was stirred vigorously for 30 minutes, then the dichloromethane layer was isolated and concentrated under reduced pressure to give a gum. The crude product was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to the desired tert -butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadia) Zol-2-yl)pyridin-2-yl)amino)propyl) carbamate (111C) was obtained. The crude sample was used in the next reaction without further purification.

Yield: 101 mg (29%).

2-methyl- N ^One -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D)

The methodology applied was similar to that described in General Method 2.

A solution of hydrogen chloride (1.2 mL, 4 M in 1,4-dioxane) was mixed with tert -butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazol-2-yl) )Pyridin-2-yl)amino)propyl)carbamate (110C) (101 mg, 0.29 mmol) was added and the mixture was stirred at room temperature for 1 h. The solvent is removed in vacuo to remove the desired 2-methyl- N ¹ -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride. (111D) was obtained as an off-white solid. The crude sample was used in the next reaction without further purification.

Yield: 93 mg (% estimated quantitation).

2-methyl- N ^One -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 203)

The methodology applied was similar to that described in General Method 3.

2-Chloro-5-methylsulfanyl-pyrimidine (100B) (51 mg, 0.32 mmol, 1.1 eq) in anhydrous N , N -dimethylformamide (2.9 mL) under nitrogen under nitrogen 2-methyl-N ¹ -(5) -(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D) (93 mg, 0.29 mmol, 1.0 eq) and cesium carbonate (283 mg, 0.87 mmol, 3.0 eq) was added to a stirred suspension. The mixture was stirred at room temperature for 16 h, heated to 40° C. for a further 16 h and then concentrated in vacuo. Water (10 mL) was added and the mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with water (10 mL) and brine (10 mL) and then dried through a phase separator. The solvent was removed in vacuo to give a yellow oil, which was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to give a crude residue which was further purified by reverse phase preparative HPLC Desired 2-methyl- N ¹ -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl) -N ³ -(5-(methylthio)pyrimidine- 2-yl)propane-1,3-diamine (Example 203) was obtained as a yellow solid.

Yield: 3.5 mg (3%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.70 (d, J = 1.8 Hz, 1H), 8.37 (s, 2H), 8.00 (dd, J = 2.3, 8.8 Hz, 1H), 6.47 (d, J = 8.1) Hz, 1H), 5.79 - 5.69 (m, 2H), 3.55 - 3.25 (m, 4H), 2.59 (s, 3H), 2.37 (s, 3H), 2.12 - 2.04 (m, 1H), 1.05 (d, J = 6.8 Hz, 3H) ; MS (ESI+) m/z 372 (M+H) ⁺ .

Scheme 17

ethyl 2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylate (112B)

In a reaction tube (R) -2-methyl- N ¹ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (544 mg, 2.19 mmol), ethyl 2- (6-bromopyridin-3-yl)cyclopropane-1-carboxylate (650 mg, 2.41 mmol), L-proline (101 mg, 0.875 mmol) potassium phosphate (928 mg, 4.37 mmol) and dimethylsulfoxide (5 mL) was added and the mixture was sparged with nitrogen for 2 min, then copper(I) iodide (83 mg, 0.437 mmol) was added. The tube was sealed under nitrogen and heated at 90° C. overnight. The reaction was cooled, diluted with ethyl acetate (20 mL), passed through a pad of celite, and the filtrate was concentrated in vacuo. The resulting residue was diluted with ethyl acetate (30 mL) and water (20 mL) and the aqueous phase was separated and extracted with ethyl acetate (2×30 mL). The combined organics were washed with water (2 x 40 mL) and then brine (2 x 50 mL), dried over magnesium sulfate and concentrated in vacuo to the crude title compound ethyl 2-(6-((( R )-2) -Methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylate (112B) was obtained as a light brown oil.

(Caution: Purification by normal phase chromatography cannot resolve several adjacent impurities).

Yield: 284 mg. ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.21 (dd, J = 2.1, 6.2 Hz, 1H), 7.95 (s, 1H), 7.26 - 7.19 (m, 1H), 7.12 (d , J = 8.1 Hz, 1H), 6.33 (d, J = 8.6 Hz, 1H), 4.22 - 4.15 (m, 2H), 3.53 - 3.46 (m, 1H), 3.39 - 3.31 (m, 2H), 3.23 - 3.17 (m, 1H), 2.49 (ddd, J = 11.8, 11.8, 11.8 Hz, 1H), 2.33 (s, 3H), 2.12 - 2.02 (m, 1H), 1.79 - 1.74 (m, 1H), 1.55 - 1.48 (m, 1H), 1.32 - 1.24 (m, 3H), 1.24 - 1.17 (m, 1H), 1.02 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 402 (M+H) ⁺ .

2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylic acid (112C)

Ethyl 2-(6-(((R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino in ethanol (6 mL) and water (4 mL) Lithium hydroxide monohydrate (143 mg, 3.41 mmol) was added to a solution of )pyridin-3-yl)cyclopropane-1-carboxylate (112B, 274 mg, 0.682 mmol), and the resulting mixture was stirred at room temperature for 1 hour. stirred for a while. The mixture was concentrated in vacuo and the resulting residue was diluted with water (8 mL). The solution was adjusted to pH about 2 with 2 M aqueous hydrochloric acid solution and then extracted with dichloromethane/methanol (20% methanol in dichloromethane, 2×10 mL). The combined organic layers were passed through a phase separator cartridge and then concentrated in vacuo to the crude title compound 2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)) Amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylic acid (112C) was obtained as a brown oil, which was used directly without further purification.

Yield: 280 mg. MS (ESI+) m / z 374 (M+H) ⁺ .

tert -Butyl 7-(2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carbonyl)-2,7-diazas Pyro[3.5]nonane-2-carboxylate (112D)

tert -Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (65 mg, 0.281 mmol) was dissolved in 2-(6-((( R )-2-methyl) in dimethylformamide (2 mL). -3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylic acid (112C, 70 mg, 0.187 mmol), 1-[bis (dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 86 mg, 0.225 mmol) and triethylamine (0.26 mL) , 1.87 mmol) and the resulting mixture was stirred at room temperature for 18 h. Upon completion the reaction mixture was concentrated in vacuo and the resulting residue was diluted with ethyl acetate (5 mL) and water (3 mL). The aqueous phase was separated and extracted with ethyl acetate (2 x 5 mL). The combined organics were washed with water (5 mL), brine (2×10 mL), dried over magnesium sulfate and then concentrated in vacuo. The obtained crude residue was purified using column chromatography (eluting with 0-10% methanol in dichloromethane) for the title compound tert -butyl 7-(2-(6-((( R )-2-methyl-) 3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane- 2-carboxylate (112D) was obtained as an off-white solid.

Yield: 69 mg. ¹H NMR (400 MHz, CDCl ₃ ) d 8.35 (s, 2H), 7.92 (d, J = 1.5 Hz, 1H), 7.15 (dd, J = 2.1, 8.5 Hz, 1H), 6.34 (d, J = 8.6) Hz, 1H), 5.96 (dd, J = 6.2, 6.2 Hz, 1H), 5.00 (s, 1H), 3.73 - 3.63 (m, 5H), 3.49 (s, 4H), 3.38 - 3.29 (m, 2H) , 3.26 - 3.18 (m, 2H), 2.37 - 2.30 (m, 4H), 2.10 - 1.98 (m, 1H), 1.89 - 1.82 (m, 1H), 1.61 - 1.53 (m, 2H), 1.44 (s, 9H), 1.29 - 1.23 (m, 2H), 1.21 - 1.14 (m, 1H), 1.02 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z (M+H) ⁺ .

(2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonane -7-yl)methanone (Example 204)

Acetic acid (0.2 mL) of trifluoroacetic dichloromethane (2 mL) of t ert- butyl-7- (2- (6 - (( (R) -2- methyl-3 - ((5- (methylthio) pyrimidine -2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate (112D, 69 mg, 0.119 mmol) was added and the mixture was stirred at room temperature for 30 min. The solvent was removed in vacuo and then azeotroped with dichloromethane (3 x 5 mL). The obtained crude residue was dissolved in dimethylsulfoxide (1.5 mL) and purified by preparative HPLC. The obtained liquid was dried under vacuum and then lyophilized from an acetonitrile/water mixture to obtain the title compound (2-(6-((( R )-2-methyl-3-((5-(methylthio)pyrimidine-) 2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonan-7-yl)methanone (204) was obtained as a foamy white solid.

Yield: 15 mg. ¹H NMR (400 MHz, CDCl ₃ ) d 8.54 (s, 1H), 8.35 (s, 2H), 7.89 - 7.86 (m, 1H), 7.16 (dd, J = 2.0, 8.6 Hz, 1H), 6.36 (d , J = 8.6 Hz, 1H), 5.98 (dd, J = 6.9, 6.9 Hz, 1H), 5.49 - 5.49 (m, 1H), 3.76 (s, 4H), 3.51 - 3.43 (m, 2H), 3.39 - 3.30 (m, 3H), 3.22 (dd, J = 6.6, 13.4 Hz, 2H), 2.38 - 2.31 (m, 4H), 2.11 - 2.01 (m, 1H), 1.89 - 1.79 (m, 5H), 1.59 - 1.52 (m, 1H), 1.27 - 1.14 (m, 1H), 1.03 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 482 (M+H) ⁺ .

Following the procedure described in Scheme 17 , the following examples were synthesized:

[Table 14]

Scheme 18

N ¹ -(5-Bromopyrazin-2-yl)-2-methyl- N 3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B )

Crude 2-methyl- N- (5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (340 mg, 1.60 mmol, 1.0 eq) in dimethylformamide (8 mL) was added to a suspension of 2,5-dibromopyrazine (457 mg, 1.92 mmol, 1.2 eq) and cesium carbonate (1.56 g, 4.80 mmol, 3.0 eq) in dimethylformamide (2 mL). The reaction mixture was heated to 90° C. for 18 hours. The solvent was removed under reduced pressure and the resulting residue was partitioned between water (20 mL) and dichloromethane (50 mL). The layers were separated and the aqueous phase was extracted with dichloromethane (3×25 mL). The combined organic phases were washed with brine (25 mL), dried through a phase separator and then concentrated to dryness in vacuo. The crude residue was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) for the title compound N ¹ -(5-bromopyrazin-2-yl)-2-methyl- N ³ -(5- (methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B) was obtained as a pale yellow gum.

Yield: 230 mg (38%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.06 (d, J = 1.3 Hz, 1H), 7.67 (d, J = 1.4 Hz, 1H), 5.67 - 5.51 (m, 2H), 3.55 - 3.31 (m, 3H), 3.25 - 3.12 (m, 1H), 2.37 (s, 3H), 2.04 (s, 1H), 1.03 (d, J = 6.9 Hz, 3H).

N ^One -(5-(2-methoxypyridin-3-yl)pyrazin-2-yl)-2-methyl- N 3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (209)

The methodology applied was similar to that described in General Method 7 (Note: 1,4-dioxane was used instead of dimethylformamide).

N ¹ -(5-Bromopyrazin-2-yl)-2-methyl- N 3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B) (50 mg, 0.135 mmol, 1.0 eq) of 2-methoxy-3-phenylboronic acid (23 mg, 0.149 mmol, 1.1 eq) in water (1 mL) and 1,4-dioxane (2 mL) under nitrogen atmosphere, To a solution of cesium carbonate (132 mg, 0.406 mmol, 3 eq) and tetrakis(triphenylphosphino)palladium(0) (15.6 mg, 0.0135 mmol, 0.1 eq). The reaction mixture was heated to 80° C. for 1 hour. The solvent was removed under reduced pressure, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3 x 15 mL). The combined organic phases were washed with brine (10 mL) and concentrated in vacuo. The obtained crude residue was purified by preparative HPLC to obtain the desired N ¹ -(5-(2-methoxypyridin-3-yl)pyrazin-2-yl)-2-methyl- N 3-(5-(methylthio). ) pyrimidin-2-yl) propane-1,3-diamine (154) was obtained as an off-white solid.

Yield: 24 mg (44%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.77 (d, J = 1.4 Hz, 1H), 8.37 (s, 2H), 8.18 - 8.13 (m, 2H), 7.99 (d, J = 1.5 Hz, 1H), 7.01 (dd, J = 4.9, 7.4 Hz, 1H), 5.69 (dd, J = 6.5, 6.5 Hz, 1H), 5.55 (dd, J = 6.1, 6.1 Hz, 1H), 4.04 (s, 3H), 3.59 - 3.48 (m, 2H), 3.42 - 3.25 (m, 2H), 2.37 (s, 3H), 2.19 - 2.06 (m, 1H), 1.06 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 398 (M+H) ⁺ .

Scheme 19

tert -Butyl (2-methyl-3-oxopropyl) carbamate (114B)

Caution - exothermic reaction; Dess-Martin periodinane (2.94 g, 6.94 mmol, 1.3 eq) was dissolved in tert -butyl (3-hydroxy-2-methylpropyl) carbamate (1.0 g, 5.34) in dichloromethane (50 mL) mmol, 1.0 eq) was added portion wise over 20 min and the mixture was stirred for 2 h in Silhorn. The mixture was diluted with dichloromethane (25 mL) and washed with 1 M aqueous sodium dithionite solution (2 x 10 mL) and saturated aqueous sodium bicarbonate solution (2 x 10 mL). The organic phase was dried through a phase separator and then concentrated in vacuo to give crude tert -butyl (2-methyl-3-oxopropyl)carbamate ( 114B), which was used immediately in the next step without further purification.

tert -Butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (114C)

The methodology applied was similar to the method described in Scheme 13 (for the production of 109D).

The crude product was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to the desired tert -butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carba Mate (114C) was obtained as a pale yellow gum.

Yield: 875 mg (47%) (MS (ESI+) m / z 345 (M+H) ⁺ .

tert-butyl(3-((2'-methoxy-[3,3'-bipyridin]-6-yl)amino)-2-methylpropyl)carbamate (114D)

The methodology applied was similar to that described in General Method 7.

The crude tert -butyl(3-((2'-methoxy-[3,3'-bipyridin]-6-yl)amino)-2-methylpropyl)carbamate (114D ) was immediately purified in the next step without further purification. was used.

Yield: 110 mg, MS (ESI+) m / z 373 (M+H) ⁺ .

N 1-(2'-methoxy-[3,3'-bipyridin]-6-yl)-2-methylpropane-1,3-diamine (114E)

The methodology applied was similar to that described in General Method 2.

Crude N ¹ -(2′-methoxy-[3,3′-bipyridin]-6-yl)-2-methylpropane-1,3-diamine (114E) was used immediately in the next step without further purification.

Crude yield: 75 mg MS (ESI+) m / z 273 (M+H) ⁺ .

N ^One -(2'-methoxy-[3,3'-bipyridin]-6-yl)-2-methyl- N 3- (5- (methylthio) pyrimidin-2-yl) propane-1,3-diamine (210)

The methodology applied was similar to that described in General Method 1.

The obtained crude residue was purified by preparative HPLC to obtain the desired product N ¹ -(2′-methoxy-[3,3′-bipyridin]-6-yl)-2-methyl- N 3-(5-( Methylthio)pyrimidin-2-yl)propane-1,3-diamine was obtained as an off-white solid (155).

Yield: 8 mg (7%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.30 (d, J = 2.1 Hz, 1H), 8.11 (dd, J = 1.9, 5.0 Hz, 1H), 7.67 (dd, J = 2.4) , 8.7 Hz, 1H), 7.57 (dd, J = 1.9, 7.3 Hz, 1H), 6.95 (dd, J = 5.0, 7.3 Hz, 1H), 6.45 (d, J = 8.4 Hz, 1H), 5.90 (dd , J = 6.3, 6.3 Hz, 1H), 5.11 (dd, J = 6.0, 6.0 Hz, 1H), 3.97 (s, 3H), 3.56 - 3.23 (m, 4H), 2.36 (s, 3H), 2.13 - 2.04 (m, 1H), 1.05 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 397 (M+H) ⁺ .

Scheme 20

tert -Butyl (3-([3,3'-bipyridin]-6-ylamino)-2-methylpropyl)carbamate (115A)

The methodology applied was similar to Scheme 19 (Example 115B) for producing the aldehyde (114B), and then a methodology similar to General Method 7 was used.

Crude N ¹ -([3,3′-bipyridin]-6-yl)-2-methylpropane-1,3-diamine (115A) was used immediately in the next step.

Yield: 253 mg (63%) ¹H NMR (400 MHz, DMSO) d 8.82 (d, J = 1.8 Hz, 1H), 8.48 (dd, J = 1.6, 4.8 Hz, 1H), 8.36 (d, J = 2.1) Hz, 1H), 8.00 - 7.96 (m, 1H), 7.76 (dd, J = 2.6, 8.7 Hz, 1H), 7.44 - 7.40 (m, 1H), 6.90 - 6.76 (m, 2H), 6.62 - 6.59 ( m, 1H), 4.10 (q, J = 5.3 Hz, 1H), 3.19 - 3.18 (m, 4H), 3.03 - 2.94 (m, 1H), 2.88 - 2.80 (m, 1H), 1.93 - 1.82 (m, 1H), 1.40 - 1.38 (m, 9H).

N ^One -([3,3'-bipyridin]-6-yl)-2-methyl-N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (211)

The methodology applied was similar to the method described in Scheme 13 (for the production of 109D).

The obtained crude residue was purified by reverse-phase chromatography (eluting with 5-95% acetonitrile in 10 mM aqueous ammonium bicarbonate solution) to obtain the desired N ¹ -([3,3′-bipyridin]-6-yl)- 2-Methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (211) was obtained as an off-white solid.

Yield: 32 mg (14%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.77 (1H, d, J = 2.1 Hz), 8.53 (1H, dd, J = 1.6, 4.8 Hz), 8.37 - 8.35 (3H, m), 7.81 - 7.77 (1H) , m), 7.63 (1H, dd, J = 2.5, 8.7 Hz), 7.33 (1H, dd, J = 4.6, 7.8 Hz), 6.50 (1H, d, J = 8.7 Hz), 5.93 (1H, dd, J = 6.1, 6.1 Hz), 5.25 (1H, dd, J = 6.1, 6.1 Hz), 3.56 - 3.25 (4H, m), 2.36 (3H, s), 2.18 - 2.00 (1H, m), 1.06 (3H) , d, J = 6.9 Hz); MS (ESI+) m / z 367 (M+H) ⁺ .

Scheme 21

2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B)

Triethylamine (49 mL, 352.25 mmol, 25 eq) was added to 2-bromo-5-iodopyridine (4.0 g, 14.09 mmol, 1.0 eq) followed by ethynyltrimethylsilane (2.9 mL, 21.13 mmol, 1.5 eq), copper(I) iodide (270 mg, 1.41 mmol, 0.1 eq), bis(triphenylphosphine)palladium(II) dichloride (99 mg, 0.14 mmol, 0.01 eq), the mixture was brought to room temperature stirred for 17 hours. The mixture was concentrated in vacuo to give a crude gum, which was purified by flash chromatography (eluting with 0-40% ethyl acetate in isohexane) to the intermediate 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) ) as a white solid.

Yield: 2.5 g (69%). MS (ESI+) m / z 254/256 (M+H) ⁺ .

2-Bromo-5-(1-(4-methoxybenzyl)-1 H -1,2,3-triazol-5-yl)pyridine (116C)

1-(azidomethyl)-4-methoxy to a solution of 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) (250 mg, 0.983 mmol, 1.0 eq) in ethanol (20 mL) Benzene (177 mg, 1.08 mmol, 1.1 eq) was added and the mixture was stirred at room temperature for 1 h. A solution of 1 M tetrabutylammonium fluoride in tetrahydrofuran (0.54 mL, 1.08 mmol, 1.1 eq) was added and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated in vacuo then purified by flash chromatography (isohexane-ethyl acetate as eluting from 0-100%) to give 2-bromo-5- (l- (4-methoxybenzyl) -1 H -1 ,2,3-triazol-5-yl)pyridine (116C) was obtained as an off-white solid.

Yield: 0.21 g (61%). MS (ESI+) m / z 347 (M+H) ⁺ .

tert -Butyl (3-((5-(1-(4-methoxybenzyl)-1 H -1,2,3-triazol-5-yl)pyridin-2-yl)amino)-2-methylpropyl)carbamate (116D)

2-Bromo-5 of dimethylsulfoxide (5 mL) (1- (4- methoxybenzyl) -1 H -1,2,3- triazol-5-yl) pyridine (116C) (135 mg, 0.391, 1.0 eq) in a degassed solution of tert -butyl (3-amino-2-methylpropyl) carbamate (96 mg, 0.508 mmol, 1.3 eq), L-proline (18 mg, 0.156 mmol, 0.4 eq), phosphoric acid Potassium (1.6.5 mg, 0.078 mmol, 0.2 eq) and copper(I) iodide (15 mg, 0.782 mmol, 0.2 eq) were added. The mixture was stirred at 90° C. for 28 h, diluted with water (20 mL) and extracted with dichloromethane (3×20 mL). The organic phases were combined and concentrated in vacuo and the crude residue was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to 2-bromo-5-(1-(4-methoxybenzyl)-1 H -1,2,3-triazol-5-yl)pyridine (116D) was obtained as a light brown solid.

Yield: 0.080 g (45%). MS (ESI+) m / z 453 (M+H) ⁺ .

tert -Butyl (3-((5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridin-2-yl)amino)-2-methylpropyl)carba Mate (116E)

The methodology applied was similar to that described in General Method 2.

Yield: 25 mg (29%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.42 (d, J = 2.0 Hz, 1H), 8.35 (s, 2H), 7.90 (dd, J = 2.4, 8.7 Hz, 1H), 7.52 (s, 1H), 6.94 - 6.90 (m, 2H), 6.44 (d, J = 9.0 Hz, 1H), 5.94 (dd, J = 6.1, 6.1 Hz, 1H), 5.49 (s, 2H), 5.09 (dd, J = 6.3, 6.3 Hz, 1H), 3.81 (s, 4H), 3.53 - 3.22 (m, 4H), 2.35 (s, 3H), 2.10 - 2.01 (m, 1H), 1.03 (d, J = 6.9 Hz, 3H), (N H is not observed).

N ^One -(5-(1) H -1,2,3-triazol-5-yl)pyridin-2-yl)-2-methyl- N 3-(4-(methylthio)phenyl)propane-1,3-diamine (212)

The methodology applied was similar to that described in General Method 3.

Yield: 18 mg (96%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.53 (d, J = 2.1 Hz, 1H), 8.38 (s, 2H), 7.84 (s, 1H), 7.82 (dd, J = 2.3, 8.7 Hz, 1H), 6.45 (d, J = 8.4 Hz, 1H), 6.17 (dd, J = 6.3, 6.3 Hz, 1H), 5.31 (dd, J = 6.0, 6.0 Hz, 1H), 3.57 - 3.26 (m, 4H), 2.36 (s, 3H), 2.19 - 2.06 (m, 1H), 1.06 (d, J = 6.8 Hz, 3H), (NH not observed); MS (ESI+) m / z 457 (M+H) ⁺ .

Scheme 22

tert -Butyl (2-methyl-3-((5-(pyridin-2-yl)pyrazin-2-yl)amino)propyl)carbamate (117B)

To a solution of 5-(pyridin-2-yl)pyrazin-2-amine (200 mg, 1.16 mmol, 1.0 eq) in dichloromethane (10 mL) tert -butyl (2-methyl-3-oxopropyl)carbamate ( 117B) (221 mg, 1.17 mmol, 1.0 eq), 3 Å molecular sieves (750 mg), acetic acid (0.266 mL, 4.65 mmol) and sodium triacetoxyborohydride (616 mg, 2.9 mmol) were added and the mixture was was stirred at room temperature for 18 hours. The reaction mixture was diluted with dichloromethane (25 mL) and filtered. The organic phase was washed with water (25 mL) and brine (25 mL) and then passed through a phase separator cartridge. The solvent was removed in vacuo to afford crude tert -butyl (2-methyl-3-((5-(pyridin-2-yl)pyrazin-2-yl)amino)propyl)carbamate (117B), which was further purified used in the next step without

Yield: 310 mg (78%).

2-methyl- N ^One -(5-(pyridin-2-yl)pyrazin-2-yl)propane-1,3-diamine (117C)

The methodology applied was similar to that described in General Method 2.

Yield: 200 mg (100%).

2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)- N 3- (5- (pyridin-2-yl) pyrazin-2-yl) propane-1,3-diamine (213)

The methodology applied was similar to that described in General Method 1.

Yield: 20 mg (10%). ¹H NMR (400 MHz, CDCl ₃ ) d 9.04 (d, J = 1.4 Hz, 1H), 8.62 - 8.61 (m, 1H), 8.37 (s, 2H), 8.10 (d, J = 8.0 Hz, 1H), 7.94 (d, J = 1.4 Hz, 1H), 7.77 - 7.72 (m, 1H), 7.20 (dd, J = 4.8, 6.5 Hz, 1H), 5.75 - 5.63 (m, 2H), 3.58 - 3.50 (m, 2H), 3.42 - 3.28 (m, 2H), 2.37 (s, 3H), 2.19 - 2.06 (m, 1H), 1.06 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 368 (M+H) ⁺ .

Following the procedure described in Scheme 22 , the following examples were synthesized:

[Table 15]

Scheme 23

5-(1-methyl-1 H -1,2,3-triazol-4-yl) pyrazin-2-amine (118B)

The methodology applied was similar to that described in General Method 7 (Note: 1,4-dioxane/water used as solvent was used instead of dimethylformamide)

Yield: 158 mg (73%) m / z 176 (M+H) ⁺ .

tert -Butyl (2-methyl-3-((5-(1-methyl-1) H -1,2,3-triazol-4-yl)pyrazin-2-yl)amino)propyl)carbamate (118C)

The methodology applied was similar to the method described in Scheme 13 (for the production of 109D).

crude tert -butyl (2-methyl-3-((5-(1-methyl- 1H -1,2,3-triazol-4-yl)pyrazin-2-yl)amino)propyl)carbamate (118C ) was directly used in the next step.

Yield: 101 mg (34%) MS (ESI+) m / z 347 (M+H) ⁺ .

2-methyl- N1 -(5-(1-methyl-1) H -1,2,3-triazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (118D)

The methodology applied was similar to that described in General Method 2.

crude 2-methyl- N ¹ -(5-(1-methyl- 1H -1,2,3-triazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (106D) directly It was used in the next step.

Yield: 71 mg (est.) MS (ESI+) m / z 367 (M+H) ⁺ .

2-methyl- N 1-(5-(1-methyl-1) H -1,2,3-triazol-4-yl)pyrazin-2-yl)- N 3- (5- (methylthio) pyrimidin-2-yl) propane-1,3-diamine (218)

The methodology applied was similar to that described in General Method 1.

Yield: 25 mg (23%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.35 (s, 3H), 8.30 (s, 1H), 8.05 (dd, J = 2.3, 8.9 Hz, 1H), 7.67 (s, 1H), 6.56 (d, J = 9.2 Hz, 1H), 6.11 - 6.06 (m, 1H), 4.14 (s, 3H), 2.63 (s, 4H), 2.35 (s, 3H), 2.19 - 2.10 (m, 1H), 1.07 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 371 (M+H) ⁺ .

Scheme 24

N ^One -(5-Bromopyrazin-2-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (119a)

The methodology applied was similar to that described in General Method 3.

Yield: 230 mg (38%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.06 (d, J = 1.3 Hz, 1H), 7.67 (d, J = 1.4 Hz, 1H), 5.67 - 5.51 (m, 2H), 5.30 (s, 2H), 3.55 - 3.31 (m, 3H), 3.25 - 3.12 (m, 1H), 2.37 (s, 3H), 2.04 (s, 1H), 1.03 (d, J = 6.9 Hz, 3H) .

2-methyl- N ^One -(5-(methylthio)pyrimidin-2-yl)- N ³ -(5-(5-(trifluoromethyl)-1 H -Pyrazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (219)

The methodology applied was similar to that described in General Method 7 (Note: 1,4-dioxane/water was used, no additional aliquots of reagent were added)

Yield: 18 mg (31%). ¹H NMR (400 MHz, DMSO) d 13.70 (s, 1H), 8.35 - 8.34 (m, 2H), 8.26 (d, J = 1.0 Hz, 1H), 8.13 - 8.12 (m, 1H), 8.00 (d, J = 1.5 Hz, 1H), 7.55 - 7.44 (m, 1H), 7.23 - 7.18 (m, 1H), 3.35 - 3.18 (m, 4H), 2.35 (s, 3H), 2.11 - 2.02 (m, 1H) , 0.95 (d, J = 6.8 Hz, 3H); MS (ESI+) m / z 426 (M+H) ⁺ .

Scheme 25

tert -Butyl (2-methyl-3-(pyrazolo[1,5- a ]pyrimidin-5-ylamino)propyl)carbamate (120A).

5-chloropyrazolo[1,5- c ]pyrimidine (500 mg, 3.26 mmol, 1 eq) and tert -butyl (3-amino-2-methylpropyl)carbamate (100E) (6.13 g, 32.56 mmol, 10 eq) was heated to 140° C. for 30 minutes under microwave irradiation. The crude reaction mixture was purified by flash chromatography (eluting with 0-100% ethyl acetate in isohexane) to tert -butyl (2-methyl-3-(pyrazolo[1,5- a ]pyrimidin-5-ylamino) ) propyl) carbamate (120A) was obtained.

Yield: 700 mg (70%). MS (ESI+) m / z 306 (M+H) ⁺ .

2-methyl- N ^One -(pyrazolo[1,5- a ]pyrimidin-5-yl)propane-1,3-diamine (120B)

The methodology applied was similar to that described in General Method 2.

MS (ESI+) m / z 229 (M+H) ⁺ .

tert -Butyl (2-methyl-3-(pyrazolo[1,5- a ]pyrimidin-5-ylamino)propyl)carbamate (220)

The methodology applied was similar to that described in General Method 3.

Yield: 20 mg (14%). ¹H NMR (400 MHz, DMSO) d 7.34 (s, 2H), 7.27 (d, J = 7.6 Hz, 1H), 6.79 (d, J = 1.8 Hz, 1H), 5.29 (d, J = 7.6 Hz, 1H) ), 5.08 (d, J = 1.5 Hz, 1H), 2.44 - 2.37 (m, 4H), 1.36 (s, 3H), 1.20 - 1.11 (m, 1H), (2 x NH not observed); MS (ESI+) m / z 330 (M+H) ⁺ .

Scheme 26

Methyl 6-bromonicotinimidate (121B)

Sodium methoxide (0.71 g, 13.22 mmol, 1.1 eq) was dissolved in an ice-cold solution of 6-bromonicotinonitrile (121A) (2.2 g, 12.02 mmol, 1.0 eq) in dioxane/water (20 mL/20 mL). was added to The mixture was stirred under ice cooling for 30 min and then allowed to warm to room temperature. After 1 h, the mixture was diluted with ethyl acetate (200 mL) and water (200 mL). The organic phase was separated and the aqueous phase was further extracted with ethyl acetate (50 mL). The organics were combined and the solvent was removed in vacuo to give crude methyl 6-bromonicotinimidate (121B), which was used in the next step without further purification.

Yield: 2.5 g (96%).

6-bromo- N '-Methylnicotinimidohydrazide (121C)

Methyl hydrazine (0.73 mL, 13.96 mmol, 1.2 eq) was added to a solution of 6-bromonicotinimidate (121B) (2.5 g, 11.63 mmol, 1.0 eq) and the mixture was stirred at room temperature for 1 h. The solvent was removed in vacuo to give crude 6-bromo- N' -methylnicotinimidohydrazide (121C), which was used in the next step without further purification.

Yield: 2 g (75%).

2-Bromo-5-(1-methyl-1 H -1,2,4-triazol-3-yl)pyridine (121D)

Formic acid (10 mL, 265 mmol, 30.4 eq) was added to 6-bromo- N '-methylnicotinimidohydrazide (121C) (2.0 g, 8.73 mmol, 1.0 eq) and the mixture was heated to reflux for 1 h. did it The reaction mixture was diluted with water (100 mL) and then extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with saturated aqueous sodium hydrogen carbonate solution (50 mL), dried over sodium sulfate and filtered. The solvent was removed in vacuo to give a crude brown residue, which was purified by flash chromatography (eluting with 0-10% methanol in dichloromethane/dichloromethane) to give 2-bromo-5-(1-methyl-1). H -1,2,4-triazol-3-yl)pyridine (121D) was obtained as an off-white solid.

Yield: 500 mg (23%). MS (ESI+) m / z 240 (M+H) ⁺ .

tert -Butyl (2-methyl-3-((5-(1-methyl-1) H -1,2,4-triazol-3-yl)pyridin-2-yl)amino)propyl)carbamate (121E)

The methodology applied was similar to that described in General Method 7 (Note: 1,4-dioxane/water was used, no additional aliquots of reagent were added)

Yield: 125 mg

2-methyl- N ^One -(5-(1-methyl-1) H -1,2,4-triazol-3-yl)pyridin-2-yl)- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (221)

The methodologies applied were similar to those described in General Method 2 and then General Method 1 for deprotection.

Yield: 25 mg (18%). ¹H NMR (400 MHz, DMSO) d 8.42 (d, J = 2.0 Hz, 1H), 8.34 (s, 2H), 8.04 (s, 1H), 7.76 (dd, J = 2.4, 8.8 Hz, 1H), 7.48 (dd, J = 6.0, 6.0 Hz, 1H), 6.82 (dd, J = 5.8, 5.8 Hz, 1H), 6.57 (d, J = 8.4 Hz, 1H), 4.16 (s, 3H), 3.32 - 3.17 ( m, 4H), 2.35 (s, 3H), 2.09 - 1.99 (m, 1H), 0.93 (d, J = 6.7 Hz, 3H); MS (ESI+) m / z 371 (M+H) ⁺ .

Scheme 27

tert -Butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (122B)

tert -Butyl N- (2-methyl-3-oxopropyl)carbamate (100E) (1.0 g, 5.34 mmol, 1.0 eq) was mixed with 5-bromopyridin-2-amine (122A) in anhydrous dichloromethane (50 mL) ) (920 mg, 5,34 mmol, 1.0 eq), acetic acid (1.2 mL, 21.36 mmol, 4.0 eq) and molecular sieves (type 4 Å, 1.0 g). The reaction mixture was stirred at room temperature for 5 min, then sodium triacetoxyborohydride (2.83 g, 13.35 mmol, 2.5 eq) was added in one portion. The mixture was stirred at room temperature for 5 hours. The reaction was quenched by careful addition of saturated aqueous sodium hydrogen carbonate solution (30 mL). The mixture was stirred vigorously for 30 min, then the dichloromethane layer was separated and washed with aqueous sodium thiosulfate solution (1 M, 15 mL). The organic phase was separated and concentrated under reduced pressure to give semi-crude tert -butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (122B) as a light brown gum, It was used in the next reaction without further purification.

Yield: 875 mg (47%).

tert -Butyl (3-((2'-methoxy-[3,3'-bipyridin]-6-yl)amino)-2-methylpropyl)carbamate (122C)

The methodology applied was similar to that described in General Method 7.

Yield: 138 mg.

N ^One -(2'-methoxy-[3,3'-bipyridin]-6-yl)-2-methyl- N ³ -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (222)

The methodologies applied were similar to those described in General Method 2 and then General Method 1 for deprotection.

Yield: 57 mg. ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.23 (d, J = 2.0 Hz, 1H), 7.68 (dd, J = 2.3, 8.7 Hz, 1H), 7.32 - 7.27 (m, 2H) ), 7.03 - 6.95 (m, 2H), 6.48 (d, J = 8.7 Hz, 1H), 5.94 - 5.90 (m, 2H), 3.82 (s, 3H), 3.56 - 3.47 (m, 1H), 3.44 - 3.35 (m, 2H), 3.28 (dd, J = 6.6, 13.4 Hz, 1H), 2.35 (s, 3H), 2.18 - 2.07 (m, 1H), 1.06 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 396 (M+H) ⁺ .

Scheme 28

6'-chloro-2 H -[1,3'-bipyridin]-2-one (123B)

2-Amino-5-iodopyridine (123A) (1.12 g, 5.00 mmol, 1.0 eq) was mixed with 2-hydroxypyridine (582 mg, 6.00 mmol, 1.2 eq), potassium carbonate in anhydrous dimethylsulfonamide (5 mL). (760 mg, 5.50 mmol, 1.1 eq), copper(I) iodide (143 mg, 0.75 mmol, 0.15 eq) and 8-hydroxyquinoline (110 mg, 0.75 mmol, 0.15 eq). The mixture was degassed under a stream of nitrogen and then heated at 130° C. for 21 h. The reaction mixture was cooled to room temperature and then poured into a mixture of 10% aqueous ammonium hydroxide solution (100 mL) and ethyl acetate (50 mL). Activated charcoal (1 g) was added and the mixture was filtered through a pad of celite, washed with ethyl acetate (2 x 50 mL). The layers were separated and the aqueous phase was extracted with ethyl acetate (2×50 mL). The combined organic extracts were washed with saturated brine (50 mL), dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude pale yellow solid was purified by flash chromatography (eluting with methanol in dichloromethane 0 to 10%) to the desired product 6 '- chloro -2 H - [1,3'- bipyridine] a-2-one as solid of mibaeksaek obtained (123B).

Yield: 245 mg, (26%). ¹H NMR (400 MHz, DMSO) d 7.88 (1H, d, J = 2.5 Hz), 7.60 (1H, ddd, J = 0.7, 2.1, 6.8 Hz), 7.49 (1H, ddd, J = 2.2, 6.7, 9.1 Hz), 7.41 (1H, dd, J = 2.7, 8.7 Hz), 6.52 (1H, dd, J = 0.4, 8.8 Hz), 6.45 (1H, ddd, J = 0.7, 1.3, 9.2 Hz), 6.28 (1H) , ddd, J = 6.7, 6.7, 1.3 Hz), 6.23 (2H, s); MS (ESI+) m/z 188 (M+H) ⁺ .

tert -Butyl (2-methyl-3-((2-oxo-2) H -[1,3'-bipyridin]-6'-yl)amino)propyl)carbamate (123C)

The methodology applied was similar to that described in Scheme 19.

6'-Chloro- 2H -[1,3'-bipyridin]-2-one (123B) was used in excess (245 mg, 1.31 mmol, 1.1 eq). A molecular sieve of 4 Å was used for the reaction. The crude product was purified by flash chromatography (dichloromethane, eluted with methanol 0-7% in) the desired product tert-butyl (2-methyl-3 - ((2-oxo -2 H - [1,3'- bipyridine ]-6′-yl)amino)propyl)carbamate (123C) was obtained as a light brown solid.

Yield: 249 mg, (58%). MS (ESI+) m/z 359 (M+H) ⁺ .

6′-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)-2 H -[1,3'-bipyridin]-2-one (223)

The methodology applied was General Method 2 using a (4:1) ratio of 4 M HCl/1,4-dioxane-water , followed by General Method 3 using cesium carbonate (3.0 eq) in combination with triethylamine (2.0 eq). similar to those described in

Yield: 152 mg (57% over 2 steps).

¹H NMR (400 MHz, DMSO) d 8.35 (2H, s), 7.91 (1H, d, J = 2.6 Hz), 7.61 (1H, ddd, J = 0.6, 2.1, 6.8 Hz), 7.52 - 7.45 (2H, m), 7.40 (1H, dd, J = 2.7, 8.9 Hz), 6.90 (1H, dd, J = 5.8, 5.8 Hz), 6.57 (1H, dd, J = 0.5, 8.9 Hz), 6.45 (1H, ddd) , J = 0.7, 1.2, 9.2 Hz), 6.28 (1H, ddd, J = 6.7, 6.7, 1.4 Hz), 3.33 - 3.14 (4H, m), 2.35 (3H, s), 2.10 - 2.00 (1H, m) ), 0.94 (3H, d, J = 6.8 Hz); MS (ESI+) m/z 383 (M+H) ⁺ .

Using the procedure described in Scheme 28 , the following examples were synthesized:

[Table 16]

Scheme 29

tert -Butyl (2-methyl-3-((5-(2-oxopyrrolidin-1-yl)pyrazin-2-yl)amino)propyl) carbamate (124B)

A solution of tert -butyl N- (3-amino-2-methylpropyl)carbamate (100E) (436 mg, 2.20 mmol, 1.1 eq) in anhydrous dimethylsulfonamide (10 mL) was dissolved in 2-bromo-5-( Pyrrolidinon-1-yl)pyrazine (124A) (510 mg, 2.00 mmol, 1.0 eq), potassium triphosphate (866 mg, 4.00 mmol, 2 eq), L-proline (94 mg, 0.80 mmol, 0.4 eq) ) and copper(I) iodide (76 mg, 0.40 mmol, 0.2 eq). The mixture was degassed, kept under a stream of nitrogen and then heated at 90° C. with stirring for 20 h. The reaction mixture was cooled to room temperature and then poured into a mixture of water (50 mL) and ethyl acetate (50 mL). The layers were separated and the aqueous phase was extracted with ethyl acetate (2×50 mL). The combined organic extracts were washed with saturated brine (50 mL), dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (eluting with 0-6% methanol in dichloromethane) to the desired product tert -butyl (2-methyl-3-((5-(2-oxopyrrolidin-1-yl)pyrazine) -2-yl)amino)propyl)carbamate (124B) was obtained as a yellow solid.

Yield: 375 mg, (54%). MS (ESI+) m/z 350 (M+H) ⁺ .

1-(5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl)pyrrolidin-2-one (233)

The methodology applied was similar to that described in General Method 3 , followed by General Method 2 using a [4:1] ratio of 4 M HCl/1,4-dioxane-water.

Yield: 71 mg (18% over 2 steps).

¹H NMR (400 MHz, CDCl ₃ ) d 9.02 (1H, d, J = 1.5 Hz), 8.35 (2H, s), 7.64 (1H, d, J = 1.5 Hz), 5.63 (1H, dd, J = 6.2) , 6.2 Hz), 5.24 (1H, dd, J = 5.7, 5.7 Hz), 3.97 (2H, t, J = 7.1 Hz), 3.55 - 3.46 (1H, m), 3.45 - 3.32 (2H, m), 3.28 - 3.19 (1H, m), 2.62 (2H, t, J = 8.1 Hz), 2.36 (3H, s), 2.20 - 2.02 (3H, m), 1.03 (3H, d, J = 6.8 Hz); MS (ESI+) m/z 374 (M+H) ⁺ .

Scheme 30

tert -Butyl 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (125B).

1,1-difluoro-2-iodo-ethane (128 mg, 0.67 mmol, 1.2 eq) was dissolved in tert -butyl 2,7-diazaspiro[3.5]nonane-7- in dimethylformamide (1.0 mL) To a suspension of carboxylate (125A) (126 mg, 0.56 mmol, 1.0 eq) and potassium carbonate (231 mg, 1.67 mmol, 3.0 eq) was added and the mixture was stirred at 60° C. for 2 h. The reaction mixture was cooled to room temperature, diluted with water and extracted with ethyl acetate. The organic phase was washed with brine, dried through a phase separator and concentrated under reduced pressure to the desired tert -butyl 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7- Carboxylate (125B) was obtained as a pale yellow oil.

Yield: 73 mg (45%). ¹H NMR (400 MHz, CDCl ₃ ) d 5.87 - 5.57 (m, 1H), 3.41 - 3.30 (m, 4H), 3.14 (s, 4H), 2.89 - 2.77 (m, 2H), 1.76 - 1.67 (m, 4H), 1.46 (s, 9H).

2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane (125C)

Trifluoroacetic acid (1.3 mL) was dissolved in dichloromethane (1.3 mL) with tert -butyl 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate ( 125B) (73 mg, 0.25 mmol, 1.0 eq) was added dropwise. The mixture was stirred for 30 min and then concentrated to dryness under reduced pressure to afford the desired product 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane (125C) as a pale yellow oil, It was used in the next reaction without further purification.

The intermediates in Table 17 were synthesized using conditions similar to those described for Intermediate 125C:

[Table 17]

Scheme 31

tert -Butyl 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (126B).

2,2,2-trifluoroethyl trifluoromethanesulfonate (153 mL, 1.06 mmol, 1.2 eq) was dissolved in tert -butyl 2,7-diazaspiro[3.5]nonane-7 in acetonitrile (2.0 mL) -Carboxylate (126A) (200 mg, 0.88 mmol, 1.0 eq), was added to a suspension of cesium carbonate (862 mg, 2.65 mmol, 3.0 eq) and the mixture was stirred at 80° C. for 16 h. The reaction mixture was cooled to room temperature, diluted with water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried through a phase separator and concentrated under reduced pressure, tert -butyl 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5] Nonane-7-carboxylate (126B) was obtained as a sticky white solid. The sample was used in the next step without further purification.

Yield: 341 mg (quantitative). ¹H NMR (400 MHz, CDCl ₃ ) d 3.38 - 3.30 (m, 4H), 3.19 (s, 4H), 3.01 (q, J = 9.4 Hz, 2H), 1.73 - 1.69 (m, 4H), 1.45 (s) , 9H).

2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane (126C)

trifluoroacetic acid (1.3 mL) in dichloromethane (1.0 mL) tert -butyl 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxyl It was added dropwise to a solution of the rate (126B) (110 mg, 0.357 mmol, 1.0 eq). The mixture was stirred for 30 min and then concentrated to dryness under reduced pressure to give 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane (126C) as a light brown oil, Estimated to be 100% yield, it was used directly in the next step without further purification.

Scheme 32

2-methyl-5-oxa-2,8-diazaspiro[3.5]nonane (127B)

Lithium aluminum hydride (1 M solution in tetrahydrofuran, 1.97 mL, 1.97 mmol, 3.0 eq) was dissolved in tert -butyl 5-oxa-2,8-diazaspiro[3.5]nonane in tetrahydrofuran (4.0 mL) under nitrogen It was added dropwise to a cooled solution of -2-carboxylate (127A) (150 mg, 0.657 mmol, 1.0 eq) at 0°C. The reaction was stirred at 0° C. for 5 min, the cooling bath was removed and the reaction heated to 70° C. for 16 h (foaming was observed at 0° C. and 35° C.). The reaction mixture was cooled to 0° C. and quenched by slow addition of water (0.075 mL), aqueous sodium hydroxide solution (15%, 0.075 mL) and then water (0.22 mL). The cooling bath was removed and the reaction mixture was stirred for 15 minutes, magnesium sulfate was added and stirred for an additional 40 minutes. The mixture was dried through a phase separator, washed with tetrahydrofuran and diethyl ether, and then concentrated to dryness under reduced pressure to obtain 2-methyl-5-oxa-2,8-diazaspiro[3.5]nonane (127B). Obtained as a light brown oil. The sample was used in the next reaction without further purification.

Yield: estimated 100%. ¹H NMR (400 MHz, CDCl ₃ ) d 3.63 - 3.56 (m, 2H), 3.47 - 3.43 (m, 2H), 3.00 (s, 2H), 2.94 - 2.86 (m, 2H), 2.83 - 2.79 (m, 2H), 2.40 (s, 3H ), N H was not observed.

The intermediates in Table 18 were synthesized using conditions similar to those described for Intermediate 127B:

[Table 18]

Scheme 33

tert -Butyl 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (128B)

The method used was similar to that described in General Method 5 using 2-(tert -butoxycarbonyl)-2-azaspiro[3.3]heptane-6-carboxylic acid to use the desired product tert -butyl 6-(dimethylcarbamoyl). )-2-azaspiro[3.3]heptane-2-carboxylate (128B) was obtained.

Yield: 111 mg (quantitative). ¹H NMR (400 MHz, DMSO) d 3.87 (s, 2H), 3.70 (s, 2H), 3.23 - 3.14 (m, 1H), 2.86 (s, 3H), 2.79 (s, 3H), 2.32 - 2.24 ( m, 4H), 1.37 - 1.36 (m, 9H).

N , N -Dimethyl-2-azaspiro[3.3]heptane-6-carboxamide (128C)

Trifluoroacetic acid (2.1 mL) was dissolved in dichloromethane (2.1 mL) in tert -butyl 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (128B) (110 mg, 0.357 mmol) , 1.0 eq) was added dropwise. The mixture was stirred for 30 min and then concentrated to dryness under reduced pressure to afford the desired product N , N -dimethyl-2-azaspiro[3.3]heptane-6-carboxamide (128C) as a light brown oil. The sample was used in the next reaction without further purification, assuming 100% yield.

Scheme 34

methyl ( S )-5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylate (129A)

The methodology applied was similar to that described in General Method 3.

Yield: 134 mg (20%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.38 (s, 2H), 8.13 (d, J = 6.9 Hz, 1H), 7.21 (d, J = 12.2 Hz, 1H), 7.12 (s, 1H), 5.62 ( t, J = 6.7 Hz, 1H), 3.93 (s, 3H), 3.64 - 3.46 (m, 2H), 3.42 - 3.22 (m, 2H), 2.38 - 2.38 (m, 3H), 2.18 - 2.09 (m, 1H), 1.07 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 422 (M+H) ⁺ .

( S )-5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxylic acid (129B)

The methodology applied was similar to that described in General Method 4.

Yield: 98 mg (quantitative).¹H NMR (400 MHz, DMSO) d 12.82 (s, 1H), 8.57 - 8.54 (m, 1H), 8.33 - 8.32 (m, 2H), 8.18 (d, J = 7.4 Hz) , 1H), 7.50 (t, J = 6.1 Hz, 1H), 7.16 (d, J = 12.5 Hz, 1H), 2.34 (s, 3H), 2.16 - 2.06 (m, 1H), 0.97 - 0.94 (m, 3H). No N H- substituting protons were observed; MS (ESI+) m / z 408 (M+H) ⁺ .

tert -Butyl ( S )-6-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate (129C)

The methodology applied was similar to that described in General Method 5.

Yield: 20 mg (54%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.27 (d, J = 1.8 Hz, 1H), 8.14 (s, 1H), 7.99 (dd, J = 1.7, 8.5 Hz, 1H), 7.51 (d, J = 8.5 Hz, 1H), 7.00 - 6.97 (m, 1H), 5.47 (dd, J = 6.7, 6.7 Hz, 1H), 4.37 (q, J = 7.2 Hz, 2H), 3.63 - 3.49 (m, 2H), 3.40 - 3.23 (m, 2H), 2.47 (s, 3H), 2.17 - 2.09 (m, 1H), 1.40 (dd, J = 7.2, 7.2 Hz, 3H), 1.06 (d, J = 6.9 Hz, 3H); MS (ESI+) m / z 489 (M+H) ⁺ .

( S )-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazol-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate salt (Example 240)

Trifluoroacetic acid (1 mL) was dissolved in anhydrous dichloromethane (1 mL) in tert -butyl ( S )-6-(5-fluoro-2-((2-methyl-3-((5-(methylthio) pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate (149) (17 mg, 0.0166 mmol, 1.0 eq) was added dropwise to a cooled solution at 0°C. The mixture was stirred for 30 min and then concentrated under reduced pressure. The obtained crude residue was purified by preparative HPLC (formic acid additive) to obtain the desired product ( S )-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidine-2) -yl)amino)propyl)amino)benzo[ d ]thiazol-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate salt (Example 189) was prepared as an off-white Obtained as a solid.

Yield: 9 mg (100%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.38 - 8.36 (m, 3H), 7.60 - 7.52 (m, 1H), 5.84 - 5.72 (m, 1H), 3.98 (d, J = 9.0 Hz, 1H), 3.84 - 3.57 (m, 6H), 3.50 - 3.24 (m, 4H), 2.86 (s, 12H), 2.38 - 2.37 (m, 3H), 2.29 - 2.13 (m, 3H), 1.09 - 1.04 (m, 3H) . No N H- substituting protons were observed; MS (ESI+) m / z 502 (M+H) ⁺ .

Using the procedure described in Scheme 34 , the following examples were prepared:

[Table 19]

Scheme 35

2-((3-((5-mercaptopyrimidin-2-yl)amino)-2-methylpropyl)amino)- N, N -dimethylbenzo [ d ]thiazole-6-carboxamide (130A)

N , N -Dimethyl-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[ d ]thiazole-6-carboxamide ( Compound 10, 75 mg, 0.18 mmol), sodium methanethiolate (126 mg, 1.80 mmol, 10.0 eq and N -methylpyrrolidine (1.5 mL) were added to a microwave vessel under nitrogen, sealed and for 1 h. Heated to 160° C. LCMS analysis showed the desired thiol 2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino) -N , It showed complete conversion to N-dimethyl benzo[ d ]thiazole-6-carboxamide (130A), which was used directly in the next step without purification.

2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino)- N, N -dimethylbenzo[ d ]thiazole-6-carboxamide (244)

Potassium hydroxide (201 mg, 3.58 mmol, 20.0 eq) and water (0.5 mL) were combined with 2-((3-((5-((difluoromethyl)thio)pyri in N-methyl pyrrolidine (1.5 mL)) Midin-2-yl)amino)-2-methylpropyl)amino) -N,N -dimethylbenzo[ d ]thiazole-6-carboxamide was added to a crude thiol solution, the mixture was cooled to -70°C and , frozen to a solid. Bromodifluoromethyldiethylphosphonate (40 uL, 0.215 mmol, 1.2 eq) was added in one portion and the reaction was allowed to warm to ambient temperature over 1 hour and then re-cooled to -70°C. An additional aliquot of bromodifluoromethyldiethylphosphonate (40 uL, 0.215 mmol, 1.2 eq) was added and the mixture was allowed to warm to ambient temperature over 1 h. Water (10 mL) was added and the mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL), then dried over magnesium sulfate. The solvent was removed in vacuo to give a crude brown oil, which was purified by flash chromatography (eluting with 0-10% methanol in dichloromethane) followed by preparative HPLC to give the desired 2-((3-((5) -((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino) -N,N -dimethylbenzo[ d ]thiazole-6-carboxamide (244) off-white was obtained as a solid of

Yield: 15 mg (19%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.42 - 8.39 (m, 2H), 7.69 (d, J = 1.4 Hz, 1H), 7.55 - 7.52 (m, 1H), 7.36 (dd, J = 1.8, 8.3 Hz) , 1H), 6.33 (s, 1H), 6.15 (t, J = 6.7 Hz, 1H), 3.65 - 3.51 (m, 2H), 3.43 - 3.29 (m, 2H), 3.08 (s, 6H), 2.20 - 2.10 (m, 1H), 1.09 - 1.06 (m, 3H); MS (ESI+) m / z 453 (M+H) ⁺ .

According to the method described in Scheme 35 , the following examples were prepared:

[Table 20]

Scheme 36

rac-tert -Butyl ((1 R ,3 R )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)carbamate (131B)

The methodology applied was similar to that described in General Method 1.

Yield: 220 mg (44%). ¹ H NMR (400 MHz, CDCl ₃ ) dd 8.35 - 8.34 (m, 4H), 5.35 (s, 1H), 5.18 - 5.15 (m, 1H), 4.84 - 4.82 (m, 1H), 4.56 - 4.53 (m , 1H), 4.36 (dd, J = 6.9, 13.8 Hz, 1H), 4.26 - 4.18 (m, 1H), 4.02 - 3.94 (m, 2H), 2.55 - 2.47 (m, 1H), 2.36 (d, J) = 1.0 Hz, 6H), 2.30 - 2.16 (m, 2H), 2.10 - 1.89 (m, 4H), 1.46 - 1.43 (m, 24H).; MS (ESI+) m / z 325 (M+H) ⁺ .

rac-(1R,3R)-N ^One -(5-(methylthio)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride (131C)

The methodology applied was similar to that described in General Method 2.

Yield: 250 mg (% quantitative). MS (ESI+) m / z 225 (M+H) ⁺ .

It was used in the next step without further purification.

rac-N , N -bis(4-methoxybenzyl)-2-(((trans)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6 -sulfonamide (131D)

The methodology applied was similar to that described in General Method 3.

Yield: 52 mg (23%). ¹H NMR (400 MHz, CDCl ₃ ) d 8.36 (s, 2H), 8.00 (d, J = 1.6 Hz, 1H), 7.74 (dd, J = 1.9, 8.5 Hz, 1H), 7.58 - 7.55 (m, 1H) ), 7.01 - 6.96 (m, 4H), 6.76 - 6.73 (m, 4H), 5.59 - 5.54 (m, 1H), 5.22 (d, J = 6.9 Hz, 1H), 4.50 - 4.44 (m, 1H), 4.35 - 4.30 (m, 1H), 4.25 (s, 4H), 3.77 - 3.76 (m, 6H), 2.37 - 2.37 (m, 6H), 2.23 - 2.07 (m, 2H); MS (ESI+) m / z 677 (M+H) ⁺ .

rac-2-((( trance )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[ d ] Thiazole-6-sulfonamide (250)

Trifluoroacetic acid (1 mL) was dissolved in rac - N , N -bis(4-methoxybenzyl)-2-((( trans )-3-((5-(methylthio) It was added dropwise to an ice-cooled solution of pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[ d ]thiazole-6-sulfonamide (131D) (45 mg, 66.48 mmol, 1.0 eq). The mixture was allowed to warm to ambient temperature over 18 h and then concentrated to dryness under reduced pressure. The obtained crude residue was purified by preparative HPLC, and the obtained clean fraction was lyophilized to the desired 2-((( trans )-3-((5-(methylthio)pyrimidin-2-yl)amino) Cyclopentyl)amino)benzo[ d ]thiazole-6-sulfonamide (Example 250) was obtained as a white powder.

Yield: 23 mg (79%). ¹ H NMR (CDCl ₃ ) d 8.36 (s, 2H), 8.17 (d, J = 1.8 Hz, 1H), 7.84 (dd, J = 2.0, 8.5 Hz, 1H), 7.59 - 7.52 (m, 1H), 5.34 (d, J = 6.5 Hz, 1H), 4.79 (s, 2H), 4.50 - 4.43 (m, 1H), 4.34 - 4.27 (m, 1H), 2.37 - 2.37 (m, 6H), 2.22 - 2.07 ( m, 2H).; MS (ESI+) m / z 437 (M+H) ⁺ .

Scheme 41

Step 1: Example 301C

To a solution of Example 301A (1 g, 3.14 mmol) in DMF (30 mL) was _{added Cs 2} CO ₃ (2.05 g, 6.28 mmol) and Example 301B (5.6 g, 62.8 mmol). The mixture was heated to 25° C. for 1 hour. TLC detected that the starting material was consumed. The reaction was filtered and the filtrate (crude Example 301C ) was used in the next step without any purification.

Step 2: Example 301D

A solution of Example 301C was treated with Boc ₂ O (1.03 g, 4.72 mmol) and stirred at room temperature for 2 h. Water (100 mL) was added, then extracted with EA (50 mL x 2), washed with water and brine _{, dried over Na 2} SO , filtered, and the filtrate was concentrated under reduced pressure, followed by silica gel chromatography ( Purification with petroleum ether/EtOAc=1/5-1/1) gave the desired product ( Example 301D , 1.2 g, yield 81%) as a yellow solid. LCMS [M+H] ⁺ = 573

Step 3: Example 301E

To a solution of Example 301D (1.2 g, 2.54 mmol) in DCM (3 mL) was added TFA (1 mL) at room temperature. After addition, the reaction mixture was stirred at room temperature for 1 h. TLC detected that the starting material was consumed, and the mixture was concentrated to give the desired product ( Example 301E 945 mg, yield: 100%) as a yellow oil, which was used in the next step without further purification.

Step 4: Example 301

To a solution of Example 301E (945 mg, 2.54 mmol) in ACN (10 mL) was added TEA (514 mg, 5.08 mmol) and Example 301F (408 mg, 2.54 mmol) at room temperature, then at 70° C. for 18 hours. added. TLC detected that the starting material was consumed. The reaction was concentrated and purified by silica gel chromatography (eluting with petroleum ether/EtOAc=3/1-5/3) to give the desired product Example 301 (780 mg, yield: 61%) as a white solid. ^{LCMS [M + H] + =} 497. 1 H NMR (400 MHz, DMSO- d 6) δ 8.78 (s, 1H), 8.34 (s, 2H), 8.12 (d, J = 1.6 Hz, 1H), 7.54 (dd, J = 8.5, 1.8 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 3.94-3.86 (m, 1H), 3.59 (dd, J = 16.6, 11.9 Hz, 6H), 3.39 ( dd, J = 19.4, 5.9 Hz, 4H), 2.85-2.81 (m, 4H), 2.33 (s, 3H).

Scheme 42

Step 1: Example 302B

A solution of Example 302A (6.77 g, 21 mmol) in THF (200 mL) under _{N 2 atmosphere was cooled to -65°C.} MeMgBr (21 mL, 63.1 mmol, 3 M in THF) was added dropwise and then stirred at -65° C. for 0.5 h. The reaction was warmed to room temperature for 2 h and quenched by addition of water (200 mL). After extraction with EtOAc (200 mL x 2), the combined organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluting with petroleum ether/EtOAc=3/1) to give the desired product Example 302B (4 g, yield 56%) as a yellow oil. LCMS [M+H] ⁺ = 339

Step 2: Example 302C

A Pd/C (400 mg) catalyst suspension in a solution of Example 302B (4 g, 0.012 mmol) in MeOH (40 mL) was introduced into the reactor. The vessel was purged with nitrogen, then hydrogen, and the reaction mixture was stirred at room temperature for 18 hours. TLC and LCMS detected that the starting material was consumed. Example 302C (4 g, yield: 100%) was obtained by filtration, concentrated and used in the next step without any purification. LCMS [M+H] ⁺ = 205

Step 3: Example 302D

To a solution of Example 301A (154 mg, 0.48 mmol) in DMF (30 mL) was _{added Cs 2} CO ₃ (313 mg, 0.96 mmol) and Example 302C (982 mg, 4.8 mmol), and the mixture was stirred for 2 h Heated to 25°C. TLC detected that the starting material was consumed. The reaction was filtered, the filtrate was concentrated under reduced pressure and purified by silica gel chromatography (eluting with petroleum ether/EtOAc=1/1 to 1/4) to give the desired product Example 302D (500 mg, yield: 100%) as a white solid was obtained as LCMS [M+H] ⁺ = 487

Step 4: Example 302E

To a solution of Example 302D (486 mg, 2 mmol) in DCM (2 mL) was added TFA (1 mL) at room temperature. After addition, the reaction mixture was stirred at room temperature for 1 h. TLC detected starting material was consumed and the mixture was concentrated to give the desired product Example 302E (532 mg, yield: 100%) as a yellow oil, which was used in the next step without further purification.

Step 5: Example 302

Example 302E (193 mg, 0.5 mmol), TEA (101 mg, 1 mmol) and Example 301F (81 mg, 0.5 mmol) were dissolved in ACN (5 mL) and the mixture was heated to 60° C. for 18 h. . LCMS detected the formation of TM. Purification by preparative HPLC gave the desired product Example 302 (15 mg, yield: 6%) as a white solid. LCMS [M+H] ⁺ = 511

¹ H NMR (400 MHz, CDCl3) δ 8.41 (s, 2H), 7.98 (s, 1H), 7.65 (s, 2H), 6.10 (s, 1H), 3.77-3.72 (m, 4H), 3.67-3.52 ( m, 4H), 3.04-2.98 (m, 4H), 2.38 (s, 3H), 1.33 (s, 3H).

Scheme 43

Step 1: Example 303B

To a solution of Example 303A (33 g, 500 mmol) and methyl carbonochloridate (49.5 g, 520 mmol) in THF (75 mL) was added KOH (56.1 g, 1000 mmol) _{in H 2 O (50 mL).} Add over 30 minutes (keep internal temperature below 40°C). After addition, the suspension was stirred at room temperature for 16 h. The suspension was filtered and the filter cake was washed with EtOH (50 mL×3) and dried in vacuo to afford the desired product Example 303B (67 g, yield 82.7%) as a white solid. LCMS [M+H] ⁺ = 163

Step 2 : Example 303C

To a solution of Example 303B (4 g, 24.52 mmol) in MeOH (300 mL) and HCl (5 mL, 4.0 M in MeOH) was added Pd/C (8 g) at room temperature. The suspension was stirred under _{15 psi of H 2} at room temperature for 44 h. LCMS showed that the starting material was consumed completely and the desired product was detected. The mixture was filtered and the filter cake was washed with MeOH (30 mL×3). The filtrate was concentrated under reduced pressure to give the product Example 303C (3.7 g, yield: 54.4%) as a yellow solid. LCMS [M+H] ⁺ = 133.2

Step 3: Example 303D

A solution of Example 303C (2.18 g, 7.84 mmol), Example 301A (0.5 g, 1.57 mmol) and DIEA (13.35 g, 103.5 mmol) in MeCN (100 mL) was heated to 80° C. and stirred for 4 h. LCMS showed that the starting material was consumed completely and the desired product was detected. Reaction mixture Example 303D was used in the next step without further purification. LCMS [M+H] ⁺ = 415.5

Step 4: Example 303E

Boc ₂ O (5 g, 22.9 mmol) was added to the solution of Example 303D at room temperature and stirred for 1 h. LCMS showed that the starting material was consumed. The reaction was diluted with DCM (200 mL), washed with water (150 mL x 2), brine (200 mL x 3), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure, which was subjected to silica gel chromatography Purification by (eluting with petroleum ether/EtOAc=2/1 to 1/9) gave the product Example 303E (680 mg, yield: 76.7% over 2 steps) as a white solid. LCMS [M+H] ⁺ = 515.5

Step 5: Example 303F

To a solution of Example 303E (840 mg, 1.63 mmol) in THF (20 mL) was _{added CaCl 2} (363 mg, 3.27 mmol) and NaBH ₄ (124 mg, 3.27 mmol) at room temperature and the reaction stirred for 16 h. did. The reaction was poured into saturated aqueous NH ₄ Cl (100 mL), diluted with EtOAc (30 mL) and separated. The aqueous layer was extracted with EtOAc (30 mL x 2), the combined organic layers were concentrated and purified by silica gel chromatography (eluting with petroleum ether/EtOAc=3/1 to 1/9) to obtain the product Example 303F (383 mg, yield: 48.3%) as a white solid. LCMS [M+H] ⁺ = 487.6

Step 6: Example 303G

To a solution of Example 303F (553 mg, 1.14 mmol) in DCM (10 mL) was added HCl/dioxane (4.0 M, 10 mL) at room temperature and the reaction stirred for 0.5 h. TLC showed that the starting material was consumed. The reaction mixture was concentrated under reduced pressure to give the product Example 303G (563 mg, yield: 100%) as a white solid. LCMS [M+H] ⁺ = 387.5

Step 7: Example 303

To a solution of Example 303G (563 mg, 1.14 mmol) and K ₂ CO ₃ (940 mg, 6.82 mmol) in DMF (10 mL) was added Example 301A (183 mg, 1.14 mmol) at room temperature. The reaction was then heated to 60° C. and stirred for 16 h. TLC showed that the starting material was consumed. The mixture was cooled to room temperature, diluted with EtOAc (30 mL) and washed with water (20 mL). The aqueous layer was extracted with EtOAc (20 mL×4), and the combined organic layers were washed with brine (20 mL×2), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel chromatography (eluting with petroleum ether/EtOAc=3/1 to 1/9) to give a crude product, which was purified by preparative HPLC to obtain product Example 303 (75.2 mg, yield: 12.93%) was obtained as a pale yellow solid. LCMS [M+H] ⁺ = 511

¹ H NMR (400 MHz, CDCl3) δ 8.37 (s, 2H), 7.96 (s, 1H), 7.66-7.58 (m, 2H), 6.82 (s, 1H), 5.87 (s, 1H), 3.83-3.73 (m, 5H), 3.59-3.44 (m, 5H), 3.01 (s, 4H), 2.38 (s, 3H), 2.14-2.08 (m, 1H).

Scheme 44

Step 1: Example 304B

Example 304A (23.6 g, 0.12 mol), NaN ₃ (15.1 g, 0.23 mol) and NH ₄ Cl (7.5 g, 0.14 mol) were suspended in DMF (250 mL), and the resulting mixture was stirred at 80° C. for 1 hour. heated to . After TLC detected that the reaction was complete, EtOAc (1 L) was added and the organic extracts were washed with water (200 mL×5), dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluting with petroleum ether/EtOAc=5/1) to give the desired product Example 304B (29 g, yield 100%) as a white solid. LCMS [M+H] ⁺ = 247

Step 2: Example 304C

To a solution of Example 304B (29 g, 0.12 mol) in EtOH (400 mL) was added HCl (22 mL) and Pd/C (1 g), and the heterogeneous mixture _{was stirred under H 2} for 18 h at room temperature. . TLC detected that most of the starting material was consumed. The mixture was filtered and the filtrate was concentrated to give the desired product Example 304C (21 g, yield: 100%) as a white solid. LCMS [M+H] ⁺ = 221

Step 3: Example 304D

To a solution of Example 304C (6.6 g, 3 mol) in DMF (50 mL) was added 2-chloro-5-(methylthio)pyrimidine (2.4 g, 15 mmol) and TEA (7.6 g, 75 mmol) . The resulting mixture was stirred at 100° C. for 1 hour. TLC detected that the starting material was consumed. The reaction mixture was concentrated and purified by silica gel chromatography (eluting with petroleum ether/EtOAc=1/1 to 1/4) to give the desired product Example 304D (450 mg, yield: 8%) as a white solid. LCMS [M+H] ⁺ = 345

Step 4: Example 304E

To a solution of Example 304D (450 mg, 1.3 mmol) in EtOH (5 mL) was added hydrazine hydrate (131 mg, 2.6 mmol) at room temperature. After 3 hours TLC detected starting material was consumed, at which point the reaction was concentrated and purified by silica gel chromatography (eluting with DCM/MeOH=10/1) to the desired product Example 304E (120 mg, yield: 43%) ) was obtained as a white solid.

Step 5: Example 304

In a solution of Example 304E (43 mg, 0.20 mmol) and 2-chloro-N,N-dimethylbenzo[d]thiazole-6-sulfonamide (37 mg, 0.13 mmol) in DMF (1 mL) DBU (40 mg, 0.26 mmol) was added. The resulting mixture was stirred at 50° C. for 1 hour. TLC detected starting material was consumed, the reaction mixture was concentrated and purified by preparative TLC to give the desired product Example 304 (30 mg, yield: 51%) as a white solid. LCMS [M+H] ⁺ = 454.9. ¹ H NMR (400 MHz, CDCl3) δ 8.35 (s, 2H), 7.99 (d, J = 1.5 Hz, 1H), 7.67 (dd, J = 8.5, 1.7 Hz, 1H), 7.59 (d, J = 8.5) Hz, 1H), 5.97 (s, 1H), 4.11 (dd, J = 9.3, 4.8 Hz, 1H), 3.74 (dd, J = 13.7, 4.3 Hz, 1H), 3.66 (s, 1H), 3.58 (dd , J = 13.6, 6.3 Hz, 2H), 2.70 (s, 6H), 2.37 (s, 3H).

Scheme 45

Step 1: Example 305

To a solution of Example 304E (30 mg, 0.14 mmol) and 2-chloro-N-methylbenzo[d]thiazole-6-sulfonamide (24.5 mg, 0.09 mmol) in DMF (2 mL) DBU (47 mg, 0.18 mmol) was added. After addition, the mixture was stirred at 50° C. for 1 h. LCMS determined that the starting material was consumed. The mixture was cooled to room temperature, diluted with EtOAc (5 mL) and washed with brine (5 mL). The organic layer was concentrated and purified by preparative TLC to give the desired product Example 305 (23.8 mg, yield: 60%) as a white solid. LCMS [M+H] ⁺ = 440.9

¹ H NMR (400 MHz, MeOD- d ₄ ) δ 8.35 (s, 2H), 8.07 (s, 1H), 7.69 (dd, J = 8.0, 1H), 7.50 (d, J = 8.0 Hz, 1H), 4.06 (m, 0.46H), 3.57 (m, 4H), 2.51 (s, 3H), 2.32 (s, 3H).

Scheme 46

Example 306 (30 mg, yield: 50%) was prepared in an analogous manner as a white solid starting from Example 306A ^{: LCMS [M+H] +} = 454.9. ¹ H NMR (400 MHz, CDCl3) δ 8.35 (s, 2H), 7.98 (d, J = 1.5 Hz, 1H), 7.66 (dd, J = 8.5, 1.7 Hz, 1H), 7.58 (d, J = 8.5) Hz, 1H), 6.04 (t, J = 5.6 Hz, 1H), 4.11 (dd, J = 9.3, 5.1 Hz, 1H), 3.74-3.64 (m, 2H), 3.61-3.54 (m, 2H), 2.70 (s, 6H), 2.36 (s, 3H).

Scheme 47

Step 1: Example 307A

To a solution of Example 306E (100 mg, 0.47 mmol) in DMF (0.5 mL) DBU (89 mg, 0.58 mmol) and methyl 2-chlorobenzo[d]thiazole-6-carboxylate (89 mg, 0.39 mmol) was added. The mixture was heated to 50° C. for 1 hour. After TLC determined that the starting material was consumed, the mixture was concentrated and purified by silica gel chromatography (eluting with petroleum ether/EtOAc=1/1 to 1/4) to the desired product Example 307A (50 mg, yield: 26%) ) as a white solid. LCMS [M+H] ⁺ = 406

Step 2: Example 307B

To a solution of Example 307A (50 mg, 0.12 mmol) in THF (1 mL) was added LiOH (0.4 mL, 1 M in water) and the resulting mixture was stirred at room temperature for 18 h. After TLC determined that the starting material was consumed, the mixture was concentrated and purified by silica gel chromatography (eluting with DCM/MeOH=10/1) to give the desired product Example 307B (5 mg, yield: 11%) as a white solid. did. LCMS [M+H] ⁺ = 392

Step 3: Example 307

To a solution of Example 307B (10 mg, 0.025 mmol) was charged dimethylamine hydrochloride (2.5 mg, 0.031 mmol), TEA (8 mg, 0.075 mmol) and HBTU (14 mg, 0.038 mmol). The resulting mixture was stirred at room temperature for 1 hour. After TLC detected that the starting material was consumed, the mixture was concentrated and purified by preparative TLC (eluting with EtOAc) to give the desired product Example 307 (2 mg, yield: 19%) as a white solid. LCMS [M+H] ⁺ = 419. ¹ H NMR (400 MHz, CDCl3) δ 8.36 (s, 2H), 7.67 (s, 1H), 7.53 (d, J = 8.2 Hz, 1H), 7.35 (dd, J = 8.3, 1.5 Hz, 1H), 5.91 (s, 1H), 4.14-4.00 (m, 1H), 3.72-3.64 (m, 2H), 3.53 (dd, J = 13.8, 6.3 Hz, 2H), 3.07 (s, 6H), 2.37 (s, 3H).

Scheme 48

Step 2: Example 308A

A Schlenk tube equipped with a magnetic stir bar was charged with TBAI (5 g, 13.6 mmol), cyclopentene (9.25 g, 136 mmol) and O-phthalimide (10 g, 68 mmol) in 250 mL of benzene. A 65% solution of TBHP (18.8 g, 136 mmol) was added before sealing the vial and the reaction mixture was stirred at 80° C. for 12 h. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate and washed with brine. The aqueous phase was extracted with ethyl acetate. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with PE/ethyl acetate=5/1) to give the desired compound Example 308A (6.5 g) as a white solid. LCMS [M+H] ⁺ = 214.

¹ H NMR (400 MHz, CDCl3) δ 2.08-2.17 (m, 1H), 2.32-2.47 (m, 1H), 2.41-2.53 (m, 1H), 2.80-2.88 (m, 1H), 5.33-5.46 ( m, 1H), 5.61-5.70 (m, 1H), 6.07-6.16 (m, 1H), 7.67-7.75 (m, 2H), 7.80-7.87 (m, 2H).

Step 2: Example 308B

To a solution of Example 308A (5 g, 23.5 mmol) in THF (25 mL) was _{added 50% hydrazine hydrate in H 2} O (3.52 g, 35.2 mmol). The mixture was stirred at 70° C. for 2 h. Then the mixture was filtered and concentrated under reduced pressure. Di-tert-butyl dicarbonate (10.2 g, 47 mmol) was added and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with PE/ethyl acetate=5/1) to give the desired compound Example 308B (550 mg) as a white solid. LCMS [M+H] ⁺ = 184.

Step 3: Example 308C

To a solution of Example 308B (700 mg, 3.825 mmol) in DCM (5 mL) was added m-CPBA (790 mg, 4.59 mmol) in small portions at 0°C. After addition, the mixture was stirred at room temperature overnight. The resulting mixture was cooled to 0° C., m-chlorobenzoic acid was filtered off and washed with additional cold DCM. The combined filtrate and washes _{were stirred with 20% NaHSO 3} for 30 min. The DCM layer was separated and extracted with 3 N NaOH (3 x 30 mL), saturated NaCl (30 mL), and then dried over _{Na 2} SO _{4 .} Evaporation left a white solid, which was purified by silica gel column chromatography (eluting with PE/ethyl acetate=5/1) to afford the desired compound Example 308C (455 mg) as a white solid. ^{LCMS [M + H] + =} 144. 1 H NMR (400 MHz, CDCl3) δ 1.07-1.17 (m, 1H), 1.48 (s, 9H), 1.66-1.76 (m, 1H), 1.89-1.96 (m , 1H), 2.05-2.16 (m, 1H), 3.41-3.50 (m, 1H), 3.53 (br s, 1H), 4.07-4.26 (m, 1H), 4.63-4.77 (m, 1H).

Step 4: Example 308D

Example 308C (445 mg, 2.28 mmol), NaN ₃ (297 mg, 4.57 mmol), NH ₄ Cl (61 mg, 1.14 mmol), 2-methoxyethanol (5 mL) and H ₂ O (1 mL) The mixture was stirred in a bath maintained at 80° C. for 16 h. The resulting solution was evaporated to dryness and the residue _{was dissolved in H 2} O (5 mL). The solution was saturated with NaCl and then extracted with DCM (4 x 5 mL). The DCM solution was evaporated and the residue was purified by silica gel column chromatography (eluting with MeOH/DCM=3%-5%) to give the desired compound Example 308D (420 mg) as a colorless oil. LCMS [M+H] ⁺ = 188.

Step 5: Example 308E

A suspension of Example 308D (420 mg, 1.74 mmol), Pd/C (cat.) in EtOH (5 mL) was _{stirred under H 2} atmosphere for 16 h at room temperature. The mixture was filtered and concentrated in vacuo. The residue was dried and used directly in the next step without further purification. Example 308E (320 mg). LCMS [M+H] ⁺ = 217.

Step 6: Example 308F

A mixture of Example 308E (150 mg, 0.694 mmol), 2-chloro-5-(methylthio)pyrimidine (111 mg, 0.694 mmol), DIPEA (180 mg, 1.4 mmol) in DMSO (5 mL) at 130 °C was stirred for 3 hours. The resulting solution was cooled to room temperature, poured into water and extracted with EtOAc (3 x 10 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with PE/EA=2/1) to give the desired compound Example 308F (100 mg) as a colorless oil. LCMS [M+H] ⁺ = 341.

¹ H NMR (400 MHz, CDCl3) δ 1.45 (s, 9H), 1.75-1.85 (m, 1H), 2.13-2.21 (m, 1H), 2.23-2.30 (m, 1H), 2.38 (s, 3H) , 3.83-4.05 (m, 3H), 5.33 (br s, 1H), 5.57 (br s, 1H), 8.35 (s, 2H).

Step 7: Example 308G

To a solution of Example 308F (100 mg, 0.294 mmol) in DCM (3 mL) was added 4 M HCl in dioxane (3 mL). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, dried and used directly in the next step without further purification. Example 308G (70.6 mg). LCMS [M+H] ⁺ = 241.

Step 8: Example 308H

Example 308G (70.6 mg, 0.256 mmol), tert-butyl 6-((2-chlorobenzo[d]thiazol-6-yl)sulfonyl)-2,6-diazaspiro[3.4 in DMF (4mL) A mixture of ]octane-2-carboxylate (130 mg, 0.294 mmol) and DIPEA (99 mg, 0.768 mmol) was stirred at 40° C. for 2 days. The resulting solution was cooled to room temperature, poured into water and extracted with EtOAc (3 x 10 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with MeOH/DCM=5%) to give the desired compound Example 308H (70 mg) as a white solid. LCMS [M+H] ⁺ = 648.

Step 9: Example 308

To a solution of Example 308H (70 mg, 0.108 mmol) in DCM (3 mL) was added 4 M HCl in dioxane (3 mL). The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified by preparative HPLC to give the title compound Example 308 (20 mg) as a white solid. LCMS [M+H] ⁺ = 548.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 1.47 - 1.56 (m, 1 H), 1.63 - 1.73 (m, 1 H), 1.97 (t, J =6.98 Hz, 2 H), 2.06 - 2.23 ( m, 2 H), 2.31 - 2.41 (m, 3 H), 3.19 (t, J =6.98 Hz, 2 H), 3.35 (s, 2 H), 3.68 (t, J =6.04 Hz, 4 H), 3.95 - 4.02 (m, 1 H), 4.15 (br. s., 1 H), 4.38 (br. s., 1 H), 7.48 (d, J =8.33 Hz, 1 H), 7.59 (dd, J) =8.33, 1.88 Hz, 2 H), 8.16 (d, J =1.88 Hz, 1 H), 8.37 (s, 2 H), 8.49 (d, J =7.79 Hz, 2 H), 8.66 (br. s. , 1 H).

Scheme 49

Step 1: Example 309A

Sodium cyclopentadienilide (a solution of 2 M in THF, 50 mL, 100 mmol, 1 equiv) was dissolved in benzylchloromethyl ether (90%, 23 g, 130 mmol, 1.3 equiv) in DMF (200 mL) at -40 °C. ) was added dropwise to the solution. After vigorous stirring at −40° C. for 20 min, the reaction mixture was poured into a 2:1 mixture of pentane/ice cold water (900 mL). After shaking to allow phase separation, the organic layer was washed twice with 150 mL of cold water _{and dried over Na 2} SO ₄ with stirring, and the temperature was kept below 0° C. to avoid isomerization of the double bond. After removal of the desiccant by filtration, the pentane was removed under vacuum at 0° C. to give (benzyloxymethyl)cyclopent-2,4-ene 1 as a pale orange oil. The resulting crude material was maintained at 0 °C under argon, diluted with THF (160 mL), cooled to -78 °C, and via cannula (-)-Ipc2BH (-)-Ipc2BH in THF (400 mL) at -78 °C at -78 °C. A solution of 1 M in THF, 100 mL, 100 mmol, 1 equiv) was added dropwise. The mixture was slowly warmed to -10 °C and stirred at that temperature for 3 days. The reaction was then quenched by the addition of MeOH (40 mL) followed by an aqueous solution of 3M of _{NaOH (40 mL) and 30% H 2} O _{2 (40 mL).} After vigorous stirring at room temperature for 24 h, THF was removed under reduced pressure and the remaining aqueous suspension was partitioned between EtOAc (400 mL) and brine (200 mL). After extraction, the organic layer _{was dried over Na 2} SO ₄ and concentrated in vacuo. The crude orange oil was purified by column chromatography (eluent: 9:1 to 8:2 heptane:EtOAc) to give Example 309A (4.8 g, 23.53 mmol, 23%) as a pale yellow oil. Rf = 0.29 (eluent: 7:3 heptane:EtOAc); 1H NMR (400 MHz, CDCl3) δ 2.23-2.32 (m, 1H), 2.35 (s, 1H), 2.63-2.73 (m, 1H), 2.81-2.88 (m, 1H), 3.28 (t, J = 8.9 Hz, 1H), 3.53 (dd, J = 5.4, 9.1 Hz, 1H), 4.29 (td, J = 4.1, 7.0 Hz, 1H), 4.52 (s, 2H), 5.53-5.58 (m, 1H), 5.70 -5.74 (m, 1H), 7.24-7.37 (m, 5H).

Step 2: Example 309B

To a solution of Example 309A (4.8 g, 23.53 mmol, 1 equiv) in anhydrous THF (100 mL) was added NaH (50% in mineral oil, 1.13 g, 28.2 mmol, 1.2 equiv) at 0° C. and the mixture was stirred at temperature Stir for 20 minutes. Benzyl bromide (BnBr, 3.6 mL, 30.5 mmol, 1.3 equiv) and tetrabutylammonium iodide (TBAI, 100 mg, 0.3 mmol, 0.01 equiv) were then added at 0° C. and the reaction mixture was stirred at room temperature. After 15 h, crushed ice was carefully added and the mixture was stirred for 30 min. After extraction with EtOAc (150 mL), the organic layer was washed with H 2 O (150 mL), brine (150 mL), dried over Na ₂ SO ₄ and concentrated in vacuo. Purification by column chromatography (eluent: 98:2 to 95:5 heptane:EtOAc) gave Example 309B (6.3 g, 21.4 mmol, 80%) as a colorless syrup. Rf = 0.41 (eluent: 9:1 heptane:EtOAc). LCMS [M+H] ⁺ =295.1H NMR (500 MHz, CDCl3) δ 2.42 (d, J = 17.4 Hz, 1H), 2.65-2.70 (m, 1H), 3.07 (brs, 1H), 3.33 and 3.44 ( AB X , J AB = 9.2 Hz, J AX = 5.7 Hz, J BX = 7.3 Hz, 2H), 4.08 (ddd, J = 3.0, 3.3, 7.0 Hz, 1H), 4.51 (d, J = 3.4 Hz, 2H) ), 4.54 (s, 2H), 5.64-5.66 (m, 1H), 5.74-5.75 (m, 1H), 7.22-7.34 (m, 10H).

Step 3: Example 309C

A 0.5 M solution of 9-BBN in THF (88 mL, 44 mmol) was added dropwise to a solution of Example 309B (6.50 g, 22.0 mmol) in anhydrous THF (10 mL) at 0° C. under nitrogen. The reaction was slowly warmed to room temperature overnight. The reaction was cooled to 0° C. and treated sequentially with EtOH (7 mL), 3 N NaOH solution (20 mL) and H ₂ O ₂ (33%, 20 mL). The resulting mixture was stirred overnight at room temperature. The resulting residue was filtered and washed with EtOAc (200 mL). To this suspension was added water (150 mL), the phases were separated and the aqueous layer was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated to dryness. The crude product was purified on silica gel (eluent: 1:1 heptane:EtOAc) to give Example 309C (6.0 g, 19.3 mmol, 87%) as a yellow oil. LCMS [M+H] ⁺ = 313.1 H NMR (400 MHz, CDCl3) δ 7.35-7.25 (m, 10H, CH-arom.), 4.52 (s, 2 H, CH2-benzyl), 4.49 (d, 1H) , J = 11.8 Hz, CHH-benzyl), 4.44 (d, 1H, J = 11.8 Hz, CHH-benzyl), 4.33-4.28 (m, 1H, H-1), 4.07 (ddd, 1H, J = 6.6 Hz) , 6.6 Hz, 4.1 Hz, H-3), 3.53 (dd, 1H, J = 9.0 Hz, 4.2 Hz, OCHH), 3.49 (d, 1H, J = 9.0 Hz, 4.3 Hz, OCHH), 2.35-2.25 ( m, 2H, H-4, H-5a), 2.05 (dddd, 1H, J = 13.5 Hz, 6.7 Hz, 3.5 Hz, 1.7 Hz, H-2a), 1.89-1.82 (m, 1H, H-2b) , 1.52-1.46 (m, 1H, H-5b).

Step 4: Example 309D

Compound Example 309C (6.0 g, 19.3 mmol) was dissolved in dry pyridine (30 mL) and cooled to 0°C. TosCl (5.5 g, 28.9 mmol) was added in small portions over 30 min. After addition, the reaction was stirred at room temperature for 18 h. The suspension was diluted with ethyl acetate (300 mL) and H ₂ O (200 mL). The organic phase was separated, washed with saturated NH4Cl solution (3 x 200 mL), brine (100 mL) and dried over MgSO4. The solvent was evaporated and the residue was purified by flash chromatography on silica gel (hexane/ethyl acetate 5:1) (1R,3S,4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl 4-Methylbenzenesulfonate (1.4 g, 3.0 mmol, 15%) was obtained as a colorless oil. Rf=0.26 (20% ethyl acetate in hexanes). LCMS [M+H] ⁺ = 467. 1H NMR (400 MHz, CDCl3) δ7.79 (m, 2H), 7.38-7.27 (m, 12H), 5.05-4.99 (m, 1H), 4.50 (s, 2H) ), 4.45 (s, 2H), 3.96-3.92 (m, 1H), 3.48-3.40 (td, 2H, J = 6.3 Hz), 2.45 (s, 3H), 2.33-2.20 (m, 2H), 2.14 2.01 (m, 2H), 1.69-1.62 (m, 1H).

The compound (1R,3S,4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl 4-methylbenzenesulfonate (1.4 g, 3.0 mmol) was dissolved in dry DMF (20 mL) and , NaN ₃ (2.1 g, 15.4 mmol) was added. The mixture was stirred at 60° C. for 14 h. After ethyl acetate (300 mL) was added, the organic layer was washed with saturated NaHCO ₃ solution (2×100 mL) and brine (100 mL) and dried over MgSO ₄ . The solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel (hexane/ethyl acetate 10:1) to give Example 309D (1.0 g, 2.95 mmol, 95%) as a colorless oil. Rf=0.54 (20% ethyl acetate in hexanes). ^{LCMS [M + H] + =} 338. 1 H NMR (400 MHz, CDCl3) δ 7.39-7.25 (m, 10H), 4.52 (dd, J = 26.0, 18.6 Hz, 2H), 4.49 (s, 2H), 3.97-3.89 (m, 1H), 3.86 (td, J = 7.0, 5.1 Hz, 1H), 3.44 (d, J = 5.5 Hz, 2H), 2.54-2.43 (m, 1 H), 2.24 (td, J) = 13.8, 6.8 Hz, 1H), 1.99 (dddd, J = 13.4, 8.7, 4.7, 1.3 Hz, 1H), 1.84 (dddd, J = 28.2, 20.8, 9.5, 4.2 Hz, 2H).

Step 5: Example 309E

Compound Example 309D (1.0 g, 2.95 mmol) was dissolved in anhydrous CH ₂ Cl ₂ (20 mL) and cooled to -78°C. A solution of 1 M BCl ₃ (40 mL) in CH ₂ Cl ₂ was added using a dropping funnel over 45 min and the mixture was stirred at -78° C. for 3 h, then allowed to warm to room temperature. The reaction was quenched with anhydrous MeOH (20 mL) at -78 °C and allowed to warm to room temperature overnight. The solvent was evaporated and the residue was purified on silica gel (hexane/ethyl acetate 1:1) to give (1S,2R,4S)-4-azido-2-(hydroxymethyl)cyclopentanol as a yellow oil ( 420 mg, 2.67 mmol, 90%) were obtained. LCMS [M+H] ⁺ =158. ¹ H NMR (400 MHz, CDCl3) δ 4.07 (dd, J = 13.3, 6.0 Hz, 1H), 4.02-3.95 (m, 1H), 3.79 (ddd, J = 10.4, 5.2, 3.0 Hz, 1H), 3.56 (dd, J = 10.4, 8.0 Hz, 1H), 2.36-2.20 (m, 4H), 1.99-1.92 (m, 1H), 1.77 (dddd, J = 14.0, 6.0, 4.5, 1.6 Hz, 1H), 1.58 (ddd, J = 13.9, 9.8, 6.8 Hz, 1H).

Compound (1S,2R,4S)-4-azido-2-(hydroxymethyl)cyclopentanol (0.42 g, 2.67 mmol) was dissolved in dry DMF (10 mL) and imidazole (198 mg, 2.94 mmol) ) was added at room temperature. After the addition of TBDPSCl (808 mg, 2.94 mmol) in small portions at 0° C., the mixture was stirred at room temperature for 16 h. The reaction _{was diluted with CH 2} Cl ₂ (200 mL), washed with saturated NH ₄ Cl (70 mL) and brine (50 mL), and dried over MgSO ₄ . The solvent was removed in vacuo and the crude product was purified by silica gel (hexane/ethyl acetate 10:1) to give Example 309E (630 mg, 1.6 mmol, 60%) as a yellow oil. LCMS [M+H] ⁺ = 360. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.69-7.66 (m, 4H), 7.49-7.40 (m, 6H), 4.15 (dd, J = 7.0, 7.0 Hz, 1H), 4.02-3.99 (m, 1H), 3.81 (dd, J = 4.8, 4.8 Hz, 1H), 3.59 (dd, J = 7.2, 7.2 Hz, 1H), 2.35-2.28 (m, 2H), 1.91 -1.88 (m, 1H), 1.83-1.78 (m, 1H), 1.70-1.62 (m, 1H), 1.08 (s, 9H).

Step 6: Example 309F

10% of Pd/C (63 mg) was added to a suspension of compound Example 309E (630 g, 1.6 mmol) in dry EtOH (100 mL). The reactants were evacuated twice to exchange an inert gas atmosphere and then connected to two balloons filled with _{H 2 .} The suspension was stirred vigorously at room temperature for 15 h. The palladium catalyst was removed using a PTFE-filter (Whatman Puradisc) and the solvent was removed under reduced pressure. The amine (588 g, 100%) was obtained as a colorless liquid and used without further purification. LCMS [M+H] ⁺ = 370.

To a solution of amine (588 mg, 1.6 mmol) in DCM (50 mL) at 0 °C was added Boc ₂ O (700 mg, 3.2 mmol) in DCM (10 mL), stirred at 0 °C, then 2 h at room temperature stirred for a while. The reaction mixture was concentrated and the crude product was purified by silica gel (hexane/ethyl acetate 5:1) to give Example 309F (650 mg, 1.38 mmol, 87%) as a colorless oil. ^{LCMS [M + H] + =} 470. 1 H NMR (400 MHz, CDCl3) δ 7.68-7.65 (m, 4H), 7.48-7.40 (m, 6H), 4.97 (br s, 1H), 4.20 (m, 1H), 4.02 (br s, 1H), 3.74 (dd, J = 4.2, 4.2 Hz, 1H), 3.51 (dd, J = 8.4, 8.4 Hz, 1H), 2.29-2.22 (m, 2H), 1.78- 1.57 (m, 3H), 1.45 (s, 9H), 1.07 (s, 9H).

Step 7: Example 309G

Compound Example 309F (630 mg, 1.34 mmol), DIEA (300 mg, 2.28 mmol) and DMAP (278 mg, 2.28 mmol) in DCM (50 mL) were added to TosCl (384 mg, 2.0 mmol) at 0 °C. . The mixture was stirred at room temperature for 14 h and then concentrated to dryness. The crude product was purified by silica gel (hexane/ethyl acetate 10:1) to give (1S,2R,4S)-4-((tert-butoxycarbonyl)amino)-2-(((tert-) Butyldiphenylsilyl)oxy)methyl)cyclopentyl 4-methylbenzenesulfonate (590 mg, 0.94 mmol, 70%) was obtained. LCMS [M+H] ⁺ = 625.

(1S,2R,4S)-4-((tert-butoxycarbonyl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl 4-methylbenzenesulfonate (590 g, 0.94 mmol) was dissolved in dry DMF (10 mL) and NaN ₃ (92 mg, 1.4 mmol) was added. The mixture was stirred at 60° C. for 14 h. After ethyl acetate (100 mL) was added, the organic layer was washed with saturated NaHCO ₃ solution (2×50 mL) and brine (50 mL) and dried over MgSO ₄ . The solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel (hexane/ethyl acetate 5:1) to tert-butyl ((1S,3R,4S)-3-azido-4-(((tert-butyldiphenylsilyl)oxy) Methyl)cyclopentyl)carbamate (460 mg, 0.93 mmol, 99%) was obtained as a colorless oil. Rf=0.54 (20% ethyl acetate in hexanes). LCMS [M+H] ⁺ = 496. ¹ H NMR (400 MHz, CDCl3) δ 7.72-7.67 (m, 4H), 7.46-7.40 (m, 6H),4.45 (br s, 1H), 4.20 (m, 1H), 4.08 (br s, 1H), 3.79-3.72 (m, 1H), 3.68-3.62 (m, 1H), 2.41-2.35 (m, 2H), 1.80-1.47 (m, 5H), 1.46 (s) , 9H), 1.08 (s, 9H).

tert-butyl ((1S,3R,4S)-3-azido-4-(((tert-butyldiphenylsilyl)oxy)methyl in 10% Pd/C (46 mg) in dry EtOH (50 mL) ) cyclopentyl) carbamate (460 g, 0.93 mmol). The reactants were evacuated twice to exchange an inert gas atmosphere and then connected to two balloons filled with _{H 2 .} The suspension was stirred vigorously at room temperature for 15 h. The palladium catalyst was removed using a PTFE-filter (Whatman Puradisc) and the solvent was removed under reduced pressure. Amine Example 309G (460 mg, 0.93 mmol, quant.) was obtained as a colorless liquid and used without further purification. LCMS [M+H] ⁺ = 470

Step 8: Example 309H

To a solution of compound Example 309G (70 mg, 0.17 mmol) in THF (5 mL) was added 1 M TBAF (1.7 mL, 0.17 mmol) _{under N 2} at 0° C. and then stirred at room temperature for 2 h. The reaction mixture was concentrated to give crude product 13, which was the next step. LCMS [M+H] ⁺ = 231.

Step 9: Example 309I

Compound Example 309H (crude, 0.17 mmol) in DMSO, 2-chloro-N,N-dimethylbenzo[d]thiazole-6-sulfonamide (46 mg, 0.17 mmol) and DIEA (65 mg, 0.51 mmol) The mixture was stirred at 60 °C overnight. The mixture was purified by preparative HPLC to give compound Example 309I (38 mg, 47%) as a white solid. LCMS [M+H] ⁺ = 472.

Step 10: Example 309J

A solution of 4 M HCl/dioxane (5 L) was added to compound Example 309I (38 mg, 0.08 mmol) in DCM (1 mL) and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to give compound Example 309J (30 mg, 100%) as a white solid. LCMS [M+H] ⁺ = 372.

Step 16: Example 309

A mixture of compound Example 309J (30 mg, 0.08 mmol), 2-chloro-5-(methylthio)pyrimidine (15 mg, 0.09 mmol) and DIEA (34 mg, 0.25 mmol) in DMSO (2.5 mL) was 120 It was stirred at ℃ for 4 hours. The mixture was purified by preparative HPLC to give compound Example 309 (5 mg, 12%) as a white solid. LCMS [M+H] ⁺ = 496.

¹ H NMR (400 MHz, CDOD) δ 8.36 (s, 2H), 8.09 (d, J = 1.6 Hz, 1H), 7.68 (dd, J = 2.0, 1.6 Hz, 1H), 7.56 (d, J = 8.4) Hz, 1H), 4.62 (br s, 5H), 4.54 (br m, 1H), 3.63-3.58 (m, 2H), 2.70 (s, 3H), 2.69-2.65 (m, 1H), 2.37 (s, 3H), 2.36-2.28 (m, 1H), 2.16-2.10 (m, 1H), 2.05-1.99 (m, 1H), 1.86-1.82 (m, 1H).

Scheme 50

Step 1: Example 310A

tert-Butyl ((1S,3S)-3-aminocyclopentyl)carbamate (1.5 g, 7.5 mmol), 2-chloro-5-(methylthio)pyrimidine (1.3 g, 8.3 mmol) was added N,N-diisopropylethylamine (3.7 mL, 22.5 mmol) at room temperature. The resulting mixture was stirred at 130° C. for 6.5 hours under _{N 2 atmosphere.} To the mixture was then added water (90 mL) and ethyl acetate (160 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography (PE:EA=20:1~PE:EA=6:1) to give Example 310A (1.8 g, yield 76%) as a yellow oil. LCMS [M+H] ⁺ = 325.

Step 2: Example 310B

To a mixture of Example 310A (1.8 g, 5.7 mmol) in methanol (2 mL) at room temperature was added HCl/dioxane (8.0 mL, 4 mol/L). The resulting mixture was stirred at room temperature under _{N 2 atmosphere for 2.5 hours.} The mixture was then evaporated to give Example 310B (1.5 g, yield 100%) as a brown solid. LCMS [M+H] ⁺ = 225.

Step 3: Example 310C

To a mixture of Example 310B (1.5 g, 5.7 mmol), ethyl 2-chlorobenzo[d]thiazole-6-carboxylate (1.4 g, 5.7 mmol) in DMSO (28 mL) DIEA (2.9 mL, 17.3 mmol) ) was added at room temperature. The resulting mixture was stirred at 80° C. under _{N 2 atmosphere for 18 hours.} To the mixture was then added water (80 mL) and ethyl acetate (150 mL). The combined organic layers were washed with brine (90 mL), dried over anhydrous sodium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography (PE:EA=30:1~5:1) to give Example 310C (1.9 g, yield 77%) as a yellow solid. LCMS [M+H] ⁺ = 430.

Step 4: Example 310D

To a mixture of Example 310C (1.9 g, 4.4 mmol) in ethanol (20 mL) and water (10 mL) at room temperature was added lithium hydroxide monohydrate (372 mg, 8.9 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 18 hours. Then, the mixture was concentrated. To the residue was added water (60 mL) and HCl (3 mol/L) until pH=5. The precipitate was filtered and the filter cake was dried to give Example 310D (1.5 g, yield 84%) as a brown solid. ^{LCMS [M + H] + =} 402. 1 H NMR (400MHz, DMSO- d 6) δ 8.49-8.47 (d, J = 6.8 Hz, 1H), 8.34 (s, 2H), 8.26-8.25 (d, J = 1.6 Hz, 1H), 7.81-7.79 (m, 1H), 7.55-7.53 (d, J = 7.2 Hz, 1H), 7.40-7.38 (d, J = 8.4 Hz, 1H), 4.39-4.33 (m, 2H), 2.35 (s, 3H), 2.21-2.19 (m, 2H), 2.12-2.10 (m, 2H), 1.98-1.94 (t, J = 13.6 Hz, 1H).

Step E: Example 310

A mixture of Example 310D (30 mg, 0.075 mmol), dimethylamine (6.75 mg, 0.15 mmol), HATU (28.5 mg, 0.075 mmol) in DCM was stirred at room temperature for 2 h. The resulting solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to give the title compound Example 310 (20 mg) as a yellow solid. ^{LCMS [M + H] + =} 429. 1 H NMR (600 MHz, DMSO- d 6) δ 1.54-1.64 (m, 2H), 1.98 (t, J = 6.75 Hz, 2H), 2.09-2.16 (m, 1H), 2.18-2.27 (m, 1H), 2.36 (s, 3H), 2.97 (s, 6H), 4.35 (br s, 2H), 7.30 (dd, J = 8.35, 1.49 Hz, 1H), 7.40 ( d, J = 8.24 Hz, 1H), 7.57 (br s, 1H), 7.74-7.81 (m, 1H), 8.35 (s, 2H), 8.58 (br s, 1H).

The following examples were synthesized using the above procedure:

[Table 31]

Scheme 51

Step 1: Example 371A

2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine in dioxane:H _{2 O=4:1 (50 mL)} (1.0 g, 4.5 mmol), 3-bromo-2-methoxypyridine (930 mg, 4.9 mmol), Pd(dppf)Cl ₂ (320 mg, 0.45 mmol) and Na ₂ CO ₃ (950 mg, 9.0 mmol) ) was stirred overnight at 110° C. under _{N 2 atmosphere.} The mixture was cooled to room temperature and water (50 mL) was added. The mixture was extracted with EtOAc (20 mL×3) and the combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography on silica gel eluting with petroleum ether:EtOAc=5:1 to give Example 371A (660 mg, 72%) as a white solid.

Step 2: Example 371

(1 S , 3 S )-N1-(5-(methylthio)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride (100 mg, 0.38 mmol) in DMSO (5 mL ), Example 371A (86 mg, 0.42 mmol) and Cs ₂ CO ₃ (376 mg, 1.15 mmol) was stirred at 130° C. for 2 days. The mixture was cooled to room temperature and water (10 mL) was added. The mixture was extracted with EtOAc (10 mL×5) and the combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by preparative HPLC to give Example 371 (10 mg, 6.4%). LCMS [M+H] ⁺ = 409.

¹ H NMR (400 MHz, CD ₃ OD) δ 8.41 (s, 2H), 8.20-8.17 (m, 2H), 8.09 (s, 1H), 7.82 (d, J = 7.2 Hz, 1H), 7.13-7.07 (m, 2H), 4.51-4.49 (m, 1H), 4.29-4.27 (m, 1H), 3.99 (s, 1H), 2.41-2.33 (m, 2H), 2.38 (s, 3H), 2.19-2.15 (m, 2H), 1.78-1.75 (m, 1H).

Scheme 52

Step 1: Example 372A

2-Fluoro-4-iodopyridin (200 mg, 0.9 mmol), pyridin-2(1H)-one (102 mg, 1.1 mmol), CuI (17 mg, 0.09 mmol), N in DMSO (5 mL) A mixture of ,N'-dimethyl-1,2-cyclohexanediamine (19 mg, 0.17 mmol) and K ₃ PO ₄ (381 mg, 1.8 mmol) _{was stirred under N 2} atmosphere at 100° C. for 3 hours. The mixture was cooled and water (20 mL) was added. The mixture was extracted with EtOAc (20 mL×3) and the combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered, concentrated and the crude product was obtained. This was purified by chromatography on silica gel eluting with petroleum ether:EtOAc = 5:1 to 1:1 to afford Example 372A (97 mg, 57%) as an off-white solid.

Step 2: Example 372

Example 372A (50 mg, 0.19 mmol), (1S,3S)-N1-(5-(methylthio)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride (40) in DMSO (3 mL) mg, 0.21 mmol) and Cs ₂ CO ₃ (185 mg, 0.57 mmol) was stirred at 130° C. for 2 days. The mixture was cooled and water (10 mL) was added. The mixture was extracted with EtOAc (10 mL×5) and the combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by preparative HPLC to give Example 372 (10 mg, 13%). ^{LCMS [M + H] + =} 395. 1 H NMR (400 MHz, CD 3 OD) δ 8.39 (s, 2H), 7.99 (d, J = 7.2 Hz, 1H), 7.70-7.64 (m, 2H), 7.20 (s, 1H), 7.05-7.03 (m, 1H), 7.67 (d, J = 9.2 Hz, 1H), 6.58-6.54 (m, 1H), 4.51-4.47 (m, 1H), 4.29-4.27 ( m, 1H), 2.42-2.33 (m, 2H), 2.38 (s, 3H), 2.20-2.14 (m, 2H), 1.78-1.74 (m, 1H).

The following examples were synthesized using the above procedure:

Scheme 54

General Method 1

6'-chloro-5-methoxy-2H-[1,3'-bipyridin]-2-one (Example 400A)

Compound 2-chloro-5-iodopyridine (191 mg, 0.8 mmol, 1.0 eq), 5-methoxypyridin-2(1H)-one (100 mg, 0.8 mmol, 1.0eq) in DMSO (5 mL), CuI (15 mg, 0.08 mmol, 0.1 eq), N,N'-dimethyl-1,2-cyclohexanediamine (23 mg, 0.16 mmol, 0.2 eq) and K ₃ PO ₄ (339 mg, 1.6 mmol, 0.2 eq) ) was stirred at 120° C. overnight in N _{2 atmosphere.} The mixture was cooled to room temperature and water (20 mL) was added. The mixture was extracted with EtOAc (20 mL×2). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered, concentrated and purified by chromatography on silica gel eluting with PE:EA = 1:1 to 400A (60 mg, 0.23 mmol). , 32%) as an off-white solid.

General Procedure 2

5-Methoxy-6′-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bi pyridin]-2-one (Example 400)

400A (53 mg, 0.22 mmol, 1.0 eq), 131C (50 mg, 0.22 mmol. 1.0 eq), tBuXphos Pd G3 (18 mg, 0.022 mmol, 0.1 eq) and tBuOK (49 mg, 0.44 mmol, 0.2 eq) was stirred overnight at 110° C. under _{N 2 atmosphere.} The mixture was cooled to room temperature and filtered. The liquid was concentrated and purified by preparative TLC followed by preparative HPLC to afford compound 400 (50 mg, 53%, TFA salt) as an off-white solid. LCMS [M+H] ⁺ = 425.

¹ H NMR (400 MHz, CD ₃ OD) δ 8.36 (s, 2H), 8.09 (s, 1H), 7.95 (d, J = 9.6 Hz, 1H), 7.50 (d, J = 9.6 Hz, 1H), 7.23 (s, 1H), 7.10 (d, J = 9.6 Hz, 1H), 6.61 (d, J = 9.6 Hz, 1H), 4.48-4.46 (m, 1H), 4.25-4.23 (m, 1H), 3.73 (s, 3H), 2.40-2.35 (m, 5H), 2.16-2.13 (m, 2H), 1.76-1.74 (m, 2H).

The following examples were synthesized using the above procedure:

Scheme 55

5-hydroxy-6′-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bi pyridin]-2-one (Example 402)

To a solution of 400 (50 mg, 0.12 mmol, 1.0 eq) in DCM (10 mL) was _{added BBr 3} (1 mL) at 0°C. The mixture was stirred at room temperature overnight, quenched with MeOH and then concentrated. The residue was purified by preparative TLC followed by preparative HPLC to give compound Example 402 (50 mg, 53%, TFA salt) as an off-white solid: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.37 ( s, 2H), 8.06 (s, 1H), 7.94 (d, J = 9.6 Hz, 1H), 7.45 (d, J = 9.6 Hz, 1H), 7.12-7.09 (m, 2H), 6.58 (d, J) = 9.6 Hz, 1H), 4.49-4.46 (m, 1H), 4.31-4.28 (m, 1H), 2.42-2.30 (m, 5H), 2.17-2.13 (m, 2H), 1.79-1.73 (m, 2H) ). [M+H] ⁺ = 411.

Scheme 56

4-Hydroxy-5'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,2'-bi pyridin]-2-one (Example 403)

From the starting material 4-(benzyloxy)pyridin-2(1H)-one, the title compound was obtained in a manner analogous to that described in General Method 1 , General Method 2 and General Method 3 ^{: 1} H NMR (400 MHz, CD _{3 )} OD) δ 8.32 (s, 2H), 7.89 (d, J = 2.2 Hz, 1H), 7.45 (d, J = 7.5 Hz, 2H), 7.39 (dd, J = 9.0, 2.7 Hz, 1H), 6.60 ( d, J = 8.6 Hz, 1H), 6.13 (dd, J = 7.5, 2.6 Hz, 1H), 5.88 (d, J = 8.6, 2.5 Hz, 1H), 4.48 - 4.25 (m, 2H), 2.35 (s) , 3H), 2.32 - 2.19 (m, 2H), 2.06 - 1.92 (m, 2H), 1.70 - 1.52 (m, 2H). [M+H] ⁺ = 411.49.

Scheme 57

General Procedure 4

6'-chloro-4-methoxy-3,3'-bipyridine (404)

(6-chloropyridin-3-yl)boronic acid (1.0 g, 6.4 mmol, 1 eq), 3-bromo-4-methoxypyridine (1.2 g, A mixture of 6.4 mmol, 1 eq), Pd(dppf)Cl ₂ (468 mg, 0.64 mmol, 0.1 eq) and Na ₂ CO ₃ (1.3 g, 12.8 mmol, 2 eq) _{was stirred under N 2} atmosphere at 105° C. overnight. did. The mixture was cooled to room temperature and quenched with water (10 mL). The mixture was extracted with EtOAc (20 mL×3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography on silica gel eluting with petroleum ether: EtOAc=5:1 to give compound 404 (200 mg, 14%) as a brown solid.

The following examples were synthesized using the above procedure:

Scheme 58

2-Chloro-5- (2, 2, 2-trifluoroethoxy) pyrimidine (408)

2,2,2-tri in a solution of 2-chloropyrimidin-5-ol (0.5 g, 3.8 mmol, 1.0 eq) and CS ₂ CO ₃ (1.49 g, 4.6 mmol, 1.2 eq) in DMF (20 mL) Fluoroethyl trifluoromethanesulfonate (0.97 g, 4.2 mmol, 1.1 eq) was added. The resulting suspension was stirred at room temperature for 16 h, then partitioned between EtOAc (30 mL) and water (80 mL). The separated aqueous layer was extracted with EtOAc (20 mL×3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated to give 408 (0.74 g), which was used directly in the next step. LCMS [M+H] ⁺ = 213.

Scheme 59

2-chloro-5- (difluoromethoxy) pyrimidine (409)

A solution of 2-chloropyrimidin-5-ol (101.6 g, 0.76 mol, 1.0 eq) in DMF (2000 mL) was charged with Cs ₂ CO ₃ (300 g, 0.92 mol, 1.2 eq), followed by 1.5 hours at room temperature stirred for a while. Sodium 2-chloro-2,2-difluoroacetate (340 g, 2.3 mol, 3.0 eq) was added and the reaction mixture was stirred at 100° C. for 3.5 h. The reaction mixture was poured into water (5 L) and extracted with EA (3 x 1 L). The combined organic phases were dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=10:1) to give 409 (70 g).

Scheme 60

2-Chloro-5-cyclopropylpyrimidine (410A)

5-bromo-2-chloropyrimidine (100 g, 518 mmol, 1.0 eq), cyclopropylboronic acid (53 g, 616 mmol, 1.2 eq) and Pd(dppf)Cl ₂ ( To a solution of 10 g, 13.7 mmol, 0.03 eq) was added Cs ₂ CO ₃ (250 g, 769 mmol, 1.5 eq) and stirred _{under N 2} at 110° C. for 12 hours. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with hexanes:ethyl acetate=15:1) to give 410A (55 g) as a yellow solid. LCMS [M+H] ⁺ =155.

General procedure 5

Tert-Butyl ((1 S , 3 S )-3-((5-cyclopropylpyrimidin-2-yl) amino) cyclopentyl) carbamate (410B)

410A (40 g, 260 mmol, 1 eq) tert-butyl ((1 S ,3 S )-3-aminocyclopentyl) carbamate (55 g, 275 mmol, 1.05 eq) and DIPEA in DMSO (400 mL) ( 105 g, 814 mmol, 3.13 eq) _{was stirred under N 2} at 110° C. for 12 h. The mixture was then cooled to room temperature and diluted with water (1000 mL). The resulting mixture was extracted with EtOAc (100 mL×3) and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography on silica gel (eluting with hexanes:ethyl acetate=5:1 to 4:1) to give compound 410B (55 g) as a pale solid. LCMS [M+H] ⁺ =319.

General procedure 6

(One S , 3 S )-N ^One -(5-cyclopropylpyrimidin-2-yl)cyclopentane-1, 3-diamine (410C)

To a solution of 410B (44 g, 13.8 mmol, 1.0 eq) in MeOH (250 mL) was added dropwise HCl (4 M in dioxane, 250 mL) and the resulting solution was stirred at room temperature for 2.5 h. After the reaction was complete, the mixture was concentrated to dryness under vacuum. The residue was redissolved in MeOH (500 mL) and the pH was adjusted to about 8 by addition of ^{ion exchange resin (Ambersep® 900 OH-form).} The mixture was filtered and the filtrate was concentrated to give 410C (44.5) as a yellow oil. LCMS [M+H] ⁺ =219.

The following examples were synthesized using the above procedure:

Scheme 61

2-Bromo-5- (methylthio) pyridine (416)

A solution of 2,5-dibromopyridine (1 g, 4.22 mmol, 1 eq) in THF (20 mL) was cooled to -78 °C under _{N 2 .} Then, n-BuLi (2.5 M, 1.77 mL, 4.43 mmol, 1.05 eq) was added dropwise at -78°C. The reaction mixture was stirred for 20 min, then 1,2-dimethyldisulfane (0.411 mL, 4.64 mmol, 1.1 eq) was added slowly. The reaction mixture was stirred for an additional 1 h and then quenched with saturated NH ₄ Cl. The reaction mixture was extracted with EA (100 mL) and water (100 mL), then washed with brine. The organic phase was dried over anhydrous Na ₂ SO ₄ . The mixture was filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE: EA = 10:1) to give 416 (86.8 mg): ESI [M+H] ⁺ = 204.1 ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.25 (t, J = 6.2 Hz, 1H), 7.41 (ddd, J = 8.9, 8.3, 1.6 Hz, 2H), 2.58 - 2.40 (m, 3H).

Scheme 62

(One S ,3 S )-N1-(thieno[3,2-b]pyridin-5-yl)cyclopentane-1,3-diamine (417)

The title compound was synthesized from the starting material 5-chlorothieno[3,2-b]pyridine using methods analogous to those described in General Method 2 and General Method 6.

The following examples were synthesized using the above procedure:

Scheme 63

General procedure 7

3- (6-chloropyridin-3-yl) pyrimidin-4 (3H) -one (419)

Pyrimidin-4(3H)-one (1.5 g, 15.61 mmol, 1.0 eq), (6-chloropyridin-3-yl)boronic acid (2.9 g, 18.73 mmol, 1.2 eq), Cu in DCM (60 mL) A solution of (OAc) ₂ (7.9 g, 43.71 mmol, 2.8 eq), pyridine (2.5 mL, 31.22 mmol, 2.0 eq) and 4 A molecular sieve (8 g) was stirred at room temperature for 48 h. The mixture was filtered through celite, the filtrate was concentrated and purified by flash column (petroleum ether: EtOAc: 1:1) to give Example 419 as a yellow solid (170 mg, 5.3% yield).

¹ H NMR (400 MHz, CDCl ₃ ) 8.43 (d, J = 2.6 Hz, 1H), 8.15 (s, 1H), 7.97 (d, J = 6.8 Hz, 1H), 7.80 (dd, J = 8.4, 2.8 Hz) , 1H), 7.53 (d, J = 8.5 Hz, 1H), 6.59 (dd, J = 6.8, 0.8 Hz, 1H); MS (ESI+) m/z 208.0 (M+H) ⁺

Using the procedure described in Scheme 63 , the following examples were prepared:

Scheme 64

Ethyl (E)-3-(2-fluoropyridin-4-yl) acrylate (423A)

To a suspension of 60% NaH (1.54 g, 38.4 mmol, 1.2 eq) in THF (15 mL)) ethyl 2-(ethoxy(propoxy)phosphoryl) acetate (8.6 g, 38.4 mmol, 1.2 eq) in THF was added dropwise at 0 °C under N _{2 atmosphere.} The mixture was stirred at 0° C. for 25 min, then a solution of 2-fluoroisonicotinaldehyde (4.0 g, 32.0 mmol, 1.0 eq) in DMF (15 mL) was added. The resulting mixture was stirred at room temperature for 16 h, then quenched with saturated aqueous NH ₄ Cl at 0 °C. The aqueous phase was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na ₂ SO ₄ and concentrated under vacuum. The residue was purified by column chromatography (PE to PE: EA = 1:15) to give 423A (3.0 g, 50% yield) as a white solid. LCMS: m/z 196 [M+H] ⁺ , rt 2.890 min. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.26 (d, J = 5.2 Hz, 1H), 7.59 (d, J = 16.1 Hz, 1H), 7.29 - 7.26 (m, 1H), 7.00 (d, J) = 1.7 Hz, 1H), 6.60 (d, J = 16.0 Hz, 1H), 4.30 (q, J = 7.1 Hz, 2H), 1.35 (t, J = 7.1 Hz, 3H).

Ethyl 3- (2-fluoropyridin-4-yl) propanoate (423B)

To a solution of 423A (3.0 g, 20.5 mmol) in EtOH (20 mL) was added 10% Pd/C (400 mg). The reaction system _{was purged with H 2} and the mixture was _{stirred under H 2} atmosphere overnight. Pd/C was filtered and the filtrate was concentrated in vacuo to give 423B (2.3 g, yield 76.7%), which was used directly: LCMS: m/z 198.1 [M+H] ⁺ , rt 2.801 min. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.12 (d, J = 5.5 Hz, 1H), 7.04 (d, J = 4.8 Hz, 1H), 6.78 (d, J = 4.0 Hz, 1H), 4.13 ( p, J = 6.8 Hz, 2H), 3.03 - 2.97 (m, 2H), 2.66 (t, J = 7.6 Hz, 2H), 1.24 (t, J = 6.7 Hz, 3H).

3- (2-oxo-1,2-dihydropyridin-4-yl) propanoic acid (423C)

To a 100 mL flask with 423B (1.53 g, 7.77 mmol) was added concentrated HCl (5 mL). The mixture was heated to 100° C. for 16 h. The mixture was then cooled to room temperature and 2 mL of water was added. The pH was adjusted to 6 by adding solid NaHCO _{3 in small portions.} The mixture was extracted with 30% i- _{PrOH in CHCl 3} and the combined organic layers were _{dried over Na 2} SO ₄ and concentrated. The residue was purified by column chromatography (MeOH/DCM) to give 423C (1.6 g, yield 91%), which was used directly in the next step. LCMS: m/z 167.8 [M+H] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.14 (d, J = 5.1 Hz, 1H), 7.05 (dt, J = 5.3, 1.7 Hz, 1H), 6.82 - 6.76 (m, 1H), 3.01 (t, J = 7.5 Hz, 2H), 2.73 (t, J = 7.5 Hz, 2H).

Methyl 3- (2-oxo-1,2-dihydropyridin-4-yl) propanoate (423D)

To a 100 mL flask with a solution of crude 423C (1.6 g) in MeOH (28 mL) was _{added 98% H 2} SO ₄ (0.15 mL). The reaction was heated to 90° C. for 16 h and then cooled to room temperature. Saturated aqueous NaCl was added to the mixture, which was extracted with 30% i _{-PrOH in CHCl 3 .} The combined organic layers were _{dried over Na 2} SO ₄ and concentrated. The residue was purified by column chromatography (MeOH:DCM = 1:15) to give the product 423D (702 mg, yield 55%). LCMS: m/z 182.1 [M+H] ⁺ .

General procedure 8

Methyl 3- (6'-fluoro-2-oxo-2H- [1,3'-bipyridin] -4-yl) propanoate (423)

423D (100.0 mg, 0.55 mmol, 1.0 eq), 2-fluoro-5-iodopyridine (147 mg, 0.66 mmol, 1.2 eq), CuI (21.0 mg, 0.10 mmol, 0.2 eq), N in a sealed tube , N'-dimethyl-1,2-cyclohexanediamine (15.5 mg, 0.10 mmol, 0.2 eq) and K ₂ CO ₃ (151.0 mg, 1.10 mmol, 2.0 eq) were added. Dioxane (3 mL) was added and the resulting mixture _{was purged with N 2} and stirred at 110° C. for 16 h. The mixture was then diluted with dichloromethane and filtered. The filtrate was washed with water and separated. The aqueous phase was extracted three times with dichloromethane. The combined organic layers were _{dried over Na 2} SO ₄ , concentrated and purified by silica gel chromatography to give 423 (42 mg, yield 27.6%). LCMS: m/z 278.08 [M+H] ⁺

Scheme 65

Methyl 6′-fluoro-2-oxo-2H-[1,3′-bipyridine]-5-carboxylate (424A)

6-fluoropyridin-3-amine (1.1 g, 10 mmol, 1.0 eq) and methyl 2-oxo-2H-pyran-5-carboxylate (1.5 g, 10 mmol, 1.0 eq) in EtOH (10 mL) The mixture was stirred at reflux overnight and then cooled to room temperature. The resulting precipitate was collected by filtration and purified by chromatography on silica gel eluting with petroleum ether: EtOAc = 2: 1 to afford compound 424A (440 mg, 18%) as an off-white solid.

Methyl 6′-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3′-bi pyridine]-5-carboxylate (424B)

From the starting materials 131C and 424A , 424B was obtained in a similar manner to that described above.

General Procedure 9

6′-(((1S, 3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3′-bipyridine ]-5-carboxylic acid (Example 424)

To a solution of compound 424B (100 mg, 0.22 mmol, 1.0 eq) in MeOH:H ₂ O = 5: 1 (10 mL) was added NaOH (35 mg, 0.88 mmol, 4.0 eq). The mixture was stirred at 50° C. for 3 h, then cooled to room temperature and acidified to pH=5-6. The resulting mixture was concentrated and redissolved in THF (50 mL). The solid was filtered and the filtrate was concentrated and triturated with EA:MeOH=5:1 to give Example 424 (90 mg, 93%) as a pale yellow solid: ¹ H NMR (400 MHz, DMSO- d _{6 )} ) δ : 8.35(s, 2H), 8.25s, 1H), 8.10 1H), 7.87 (d, J = 9.2 Hz, 1H), 7.56 (s, 1H), 6.87 (s, 1H), 6.51 (d, J = 9.2 Hz, 1H), 4.36-4.33 (m, 2H), 2.35 (s, 3H), 2.22-2.10 (m, 2H), 2.01-1.93 (m, 2H), 1.57-1.55 (m, 2H) .

The following examples were synthesized using the above procedure:

Scheme 66

2-chloro-5-((trimethylsilyl)ethynyl)pyridine (431A)

To a degassed solution of 2-chloro-5-iodopyridine (5 g, 20.88 mmol, 1.0 eq) in triethylamine (35 mL) ethynyltrimethylsilane (3.2 mL, 22.97 mmol, 1.1 eq), CuI (397.7 mg) , 2.09 mmol, 0.1 eq) and Pd(PPh ₃ ) ₂ Cl ₂ (1.5 g, 2.09 mmol, 0.1 eq) were added. The reaction mixture was stirred at room temperature under nitrogen for 16 h. Water (150 mL) was added and the system _{was extracted with Et 2} O (100 mL×2). The combined organic layers were _{dried over Na 2} SO ₄ , filtered and concentrated under reduced pressure. The resulting crude 431A (6.2 g, black solid) was used in the next step without further purification. MS (ESI+) m/z 209.9 (M+H) ⁺

2-Chloro-5-ethynylpyridine (431B)

A solution of 431A (crude, 20.88 mmol, 1.0 eq) and K ₂ CO ₃ (2.9 g, 20.88 mmol, 1.0 eq) in methanol (50 mL) was stirred at room temperature for 2 h. After removal of the solvent under reduced pressure, DCM (150 mL) was added and the mixture was filtered. The filtrate was concentrated and purified by flash column (petroleum ether: EtOAc = 10:1) to give compound 432B as a yellow solid (1.0 g, 34.8% yield in 2 steps). MS (ESI+) m/z 138.1 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ): 8.55 (d, J = 2.1 Hz, 1H), 7.98 (dd, J = 8.4, 2.4 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 4.56 (s, 1H).

2-chloro-5- (1- ((trimethylsilyl) methyl) -1H-1,2,3-triazol-4-yl) pyridine (431C)

431B (345 mg, 2.54 mmol, 1.0 eq), (azidomethyl)trimethylsilane (327 mg, 2.54 mmol,1.0 eq), CuI (48 mg, 0.25 mmol, 0.1eq), NEt _{3 in} THF (10 mL) (513 mg, 5.08 mmol, 2.0 eq) was stirred at room temperature for 16 h. The reaction mixture was then concentrated to give 431C (673 mg), which was used without further purification.

2-chloro-5- (1-methyl-1H-1,2,3-triazol-4-yl) pyridine (431)

To a solution of 431C in THF (10 mL) was added TBAF (0.80 g, 3.0 mmol, 1.2 eq) and the resulting solution was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography on silica gel (PE:EA=1:1) to give 431 (150 mg).

Scheme 67

2-chloro-5- (iodoethynyl) pyridine (432A)

LDA (4.4 mL, 8.73 mmol, 1.2 eq) was added dropwise to a solution of 431B (1.0 g, 7.27 mmol, 1.0 eq) in THF (15 mL) at -78°C under nitrogen. The mixture was stirred at -78 °C for 0.5 h, then a solution of iodine (2.0 g, 8.00 mmol, 1.1 eq) in THF (10 mL) was added dropwise. The resulting solution was slowly warmed to room temperature and stirred for an additional 5 h, then quenched by addition of saturated ammonium chloride solution (25 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc (50 mL). The combined organic layers were washed with Na ₂ S ₂ O ₃ (25 mLx2) and brine (30 mL), dried over Na ₂ SO ₄ and concentrated under reduced pressure. The resulting residue was purified by flash column (petroleum ether: EtOAc: 20:1) to give 432A as a yellow solid (1.56 g, 82.1% yield). MS (ESI+) m/z 263.9 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ): 8.51 (d, J = 2.0 Hz, 1H), 7.94 (dd, J = 8.3, 2.4 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H).

2-chloro-5- (5-iodo-1-((trimethylsilyl) methyl) -1H-1,2,3-triazol-4-yl) pyridine (432B)

CuI (108.4 mg, 5.69 mmol, 1.0 eq) and Et ₃ N (1.6 mL, 11.39 mmol, 2.0 eq) were stirred in THF (60 mL) under nitrogen at room temperature for 1 h. A solution of 432A (1.5 g, 5.69 mmol, 1.0 eq) and (azidomethyl)trimethylsilane (735.8 mg, 5.69 mmol, 1.0 eq) in THF (20 mL) was added to the above catalyst solution in one portion. The mixture was then stirred at room temperature for 16 h. The reaction was quenched by addition of 10% ammonium chloride solution (15 mL) and concentrated. The residue was washed with water (30 mL) and EtOAc (8 mL) to give 432B as a yellow solid (1.6 g, 72.7% yield), which was used directly in the next step. ¹ H NMR (400 MHz, DMSO- d ₆ ) 8.74 (d, J = 2.3 Hz, 1H), 8.14 (dd, J = 8.3, 2.5 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 3.80 (s, 2H), 0.00 (s, 9H). MS (ESI+) m/z 393.0 (M+H) ⁺

2-chloro-5- (5-iodo-1-methyl-1H-1,2,3-triazol-4-yl) pyridine (432C)

To a solution of 432B (1.6 g, 4.07 mmol, 1.0 eq) in THF (70 mL) was added water (0.15 mL, 8.15 mmol, 2.0 eq) followed by TBAF (4.9 mL, 4.89 mmol, 1.2 eq) at 0 °C was added dropwise. The resulting reaction mixture was stirred at 0° C. for 15 min and poured into water (100 mL), which was extracted with DCM (300 mL). The separated organic layer was washed with brine (80 mL), dried over Na ₂ SO ₄ and concentrated. The residue was purified by flash column (petroleum ether: EtOAc: DCM: 2: 1 :1) to give 432B as a yellow solid (810 mg, 62.3% yield). MS (ESI+) m/z 320.9 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ): 8.91 (d, J = 2.1 Hz, 1H), 8.32 (dd, J = 8.4, 2.5 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 4.13 (s, 3H).

2-chloro-5- (5-fluoro-1-methyl-1H-1,2,3-triazol-4-yl) pyridine (432)

A suspension of 406B (800 mg, 2.50 mmol, 1.0 eq) and KF (1.5 g, 25.00 mmol, 10.0 eq) in acetonitrile/water (14 mL, 1:1) was reacted in a microwave reactor at 160° C. for 20 min. . After evaporation under reduced pressure, the residue was dissolved with DCM (300 mL) and filtered. The filtrate was concentrated and purified by flash column (petroleum ether: EtOAc: DCM: 2: 1 :1) to give Example 432 as a yellow solid (220 mg, 41.4% yield). MS (ESI+) m/z 213.0 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) d 8.77 (d, J = 2.4 Hz, 1H), 8.17 (dd, J = 8.3, 2.4 Hz, 1H), 7.67 (d, J = 8.3 Hz, 1H) , 4.01 (s, 3H).

Scheme 68

4-nitrophenyl (6-bromopyridin-3-yl) carbamate (433A)

To a solution of 6-bromopyridin-3-amine (600 mg, 3.47 mmol, 1.0 eq) in acetonitrile (20 mL), 4-nitrophenyl carbonochloridate (768.9 mg, 3.81 mmol, 4 mL of acetonitrile) heavy, 1.1 eq) was added dropwise and the system temperature was maintained below 40°C. After addition, the mixture was stirred at room temperature for 30 min and a yellow precipitate was observed. The precipitate was filtered and washed with acetonitrile (2 mL) to give 433A as a yellow solid (1.1 g, about 50% purity), which was used in the next step without further purification.

General procedure 10

3-(6-bromopyridin-3-yl)-1-methylimidazolidine-2,4-dione (Example 433)

A solution of methyl 2-(methylamino)acetate hydrochloride (454.1 mg, 3.25 mmol, 1.0 eq) and DIPEA (1.7 mL, 9.76 mmol, 3.0 eq) in acetonitrile (15 mL) was stirred at room temperature for 15 min. 433A (1.1 g, 3.25 mmol, 1.0 eq) was added and the resulting system stirring was continued at room temperature for 10 min. The mixture was concentrated and the residue was purified by flash column (petroleum ether: EtOAc: 1:1) to give compound example 433 as a yellow oil (510 mg, 54.6% yield by 2 steps). MS (ESI+) m/z 270.1 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) d 8.48 - 8.52 (m, 1H), 7.82 - 7.89 (m, 2H), 4.19 (s, 2H), 3.00 (s, 3H).

Scheme 69

Methyl (2-methoxyethyl) glycinate (434A)

To a solution of 2-methoxyethan-1-amine (2.9 mL, 33.36 mmol, 1.0 eq) in THF (40 mL) was added Et ₃ N (9.3 mL, 66.90 mmol, 2.0 eq) dropwise followed by methyl 2-bro Parent acetate (2.8 mL, 29.58 mmol, 0.9 eq) was added. The reaction mixture was stirred at room temperature for 19 hours. The reaction mixture was diluted with EA, then washed with water and brine. The organic phase was _{dried over Na 2} SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 20:1) to give 434A (830 mg) as a colorless liquid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 3.62 (s, 3H), 3.38-3.34 (m, 4H), 3.23 (s, 3H), 2.65 (t, J = 5.6 Hz, 2H).

3-(6-bromopyridin-3-yl)-1-(2-methoxyethyl)imidazolidine-2,4-dione (Example 434)

Example 434 was obtained as a yellow solid from starting materials 434A and 433A in a manner analogous to that described above ^{: 1} H NMR (400 MHz, CDCl3) δ 8.59 (d, J = 2.6 Hz, 1H), 7.74 (dd, J) = 8.5, 2.7 Hz, 1H), 7.58 (d, J = 8.5 Hz, 1H), 4.22 (s, 2H), 3.70 - 3.64 (m, 2H), 3.64 - 3.60 (m, 2H), 3.39 (s, 3H). ESI (M+H) ⁺ = 314.3.

Scheme 70

1- (4-iodophenyl) pyridin-2 (1H) -one (435)

1,4-diiodobenzene (1.0 g, 3.0 mmol, 1.0 eq), pyridin-2(1H)-one (288 mg, 3 mmol, 1.0 eq), CuI (58 mg, 0.3) in DMSO (10 mL) mmol, 0.1 eq) and K ₂ CO ₃ (828 mg, 6 mmol, 2.0 eq) _{was stirred under N 2} at 130° C. for 2 h. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (30 mL). The organic mixture was washed sequentially with water and brine, _{dried over Na 2} SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 10:1 to EA) to give 200 mg of Example 435 as a white solid. ESI (M+H) ⁺ = 298.09

Scheme 71

Methyl 3- (2-chloropyridin-4-yl) -2,2-dimethylpropanoate (436A)

A solution of methyl isobutyrate (3.3 g, 32.0 mmol, 2.08 eq) in THF (25 mL) was dissolved in LDA (17 mL, 34.0 mmol, 2.2 eq) in THF (50 mL) _{at -78 °C under N 2 atmosphere over 15 min.} ) was added dropwise to the solution. The resulting mixture was stirred at -78 °C for 45 min, then treated with a solution of 2-chloro-4-(chloromethyl)pyridine (2.5 g, 15.4 mmol, 1.0 eq) in THF (6 mL) over 5 min. did. The cooling bath was removed and the reaction mixture was stirred at room temperature for 18 hours. 1.0 N aqueous hydrochloric acid was added dropwise to the solution (50 mL) to quench the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic layers were washed with water, dried over sodium sulfate and concentrated in vacuo. The crude product was purified by column chromatography to give compound 436A (3.2 g) as a yellow oil: LCMS: m/z 228.0. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.33 - 8.21 (m, 1H), 7.16 - 7.07 (m, 1H), 6.98 (dd, J = 5.1, 1.5 Hz, 1H), 3.68 (s, 3H), 2.84 (s, 2H), 1.21 (s, 6H).

Methyl 2,2-dimethyl-3- (2-oxo-1,2-dihydropyridin-4-yl) propanoate (436B)

A solution of 436A (1.2 g, 5.3 mmol, 1.0 eq), sodium acetate (868 mg, 10.6 mmol, 2.0 eq) in acetic acid (5.3 mL) was heated in a microwave reactor at 160° C. for 1 h. The mixture was concentrated in vacuo and the residue was poured into water. The aqueous phase was extracted twice with 15% isopropanol in DCM. The combined organic layers were washed with saturated aqueous NaHCO ₃ , dried over Na ₂ SO ₄ and concentrated in vacuo. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 20: 1) to give 436B (330 mg) as a white solid: ESI [M+H] ⁺ =210.24.

General Procedure 11

Methyl 3-(6'-chloro-2-oxo-2H-[1,3'-bipyridin]-4-yl)-2,2-dimethylpropanoate (436C)

436B (330 mg, 1.58 mmol, 1.0 eq), 2-chloro-5-iodopyridine (567 mg, 2.37 mmol, 1.5 eq), N 1 ,N ₂ -dimethylcyclohexane- _{1 in} dioxane (8 mL) A suspension of ,2-diamine (44.8 mg, 0.316 mmol, 0.2 eq), CuI (60 mg, 0.316 mmol, 0.2 eq) and K ₃ CO ₃ (436 mg, 3.16 mmol, 2.0 eq) _{under N 2} overnight at 110° C. stirred in. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE: EtOAc = 3:1 to PE: EtOAc = 1:1) to give 436C (350 mg) as a yellow oil: ESI [M+H] ⁺ =321.77 .

Scheme 72

(E)-4-((dimethylamino)methylene)isochroman-1,3-dione (437A)

To a solution of 2-(carboxymethyl)benzoic acid (10 g, 50 mmol, 1.0 eq) in DMF (100 mL) at 0° C. was added phosphoryl chloride (10 mL, 107 mmol, 2.1 eq) with stirring. The resulting mixture was stirred for an additional hour and then poured into ice water. The precipitate formed was collected by filtration and washed with water to give 437A as a yellow solid (10 g).

Methyl 1-oxo-1H-isochromene-4-carboxylate (437B)

Dry hydrogen chloride gas was passed through a stirred solution of 437A (6.2 g, 0.03 mmol, 1.0 eq) in methanol (180 mL) at room temperature for 2 h. The solution was heated at reflux for 2 h and then concentrated under reduced pressure. The residue was diluted with water and then extracted with DCM (20 mLx3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by column chromatography on silica gel (PE:EA=3:1) to give 437B (1.9 g).

Methyl 2- (6-chloropyridin-3-yl) -1-oxo-1,2-dihydroisoquinoline-4-carboxylate (Example 437)

A solution of 437B (0.83 g, 0.41 mmol, 1.0 eq) and 6-chloropyridin-3-amine (0.53 g, 0.41 mmol, 1.0 eq) in AcOH (15 mL) was heated to 120° C. and stirred for 2 h. The system was cooled to room temperature and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (PE:EA=10:1) to give Example 437 (400 mg).

Scheme 73

General Procedure 12

2- (6-chloropyridin-3-yl) pyridazin-3 (2H) -one (438)

2-Chloro-5-iodopyridine (5.95 g, 25 mmol, 1.0 eq), pyridazin-3(2H)-one (2.52 g, 26.3 mmol, 1.05 eq), CuI (475 mg) in DMSO (25 mL) , 2.5 mmol, 0.1 eq), trans-N,N'-dimethyl-1,2-cyclohexanediamine (534 mg, 3.76 mmol, 0.15 eq) and K ₂ CO ₃ (6.9 g, 50 mmol, 2.0 eq) of The mixture was stirred at 120° C. overnight under _{N 2 atmosphere.} The mixture was cooled to room temperature and filtered. The filtrate was diluted with water and then extracted with EA (200 mL x 2). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The resulting residue was purified by chromatography on silica gel eluting with PE:EA=5:1 to 1:1 to give 438 (3.6 g, 69%) as a white solid.

Scheme 74

Methyl 2- (6-chloropyridin-3-yl) -3-cyanopropanoate (439A)

LiHMDS (24.24 mL, 24.24 mmol, 1.5 eq) in a cold (-78 °C) solution of methyl 2-(6-chloropyridin-3-yl) acetate (3.0 g, 16.2 mmol, 1.0 eq) in THF (30 mL) was added dropwise. The reaction mixture was stirred at -78 °C for 2 h. 2-Bromoacetonitrile (1.7 mL, 24.24 mmol, 1.5 eq) was added dropwise at -78°C. The reaction mixture was stirred at −78° C. for an additional 2 h and then quenched with water. The reaction mixture was extracted 3 times with EA. The combined organic phases were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 2:1) to give 439A (1.46 g) as a yellow oil. ESI (M+H) ⁺ = 225.3

3- (6-chloropyridin-3-yl) pyrrolidin-2-one (439B)

_{NaBH 4} (590 mg, 15.6 ) in a cold (0 °C) solution of 439A (700 mg, 3.1 mmol, 1.0 eq) and CoCl ₂ (370 mg, 1.56 mmol, 0.5 eq) in THF/water (6 mL/3 mL) mmol, 5.0 eq) was added at 0° C. under _{N 2 .} The reaction mixture was stirred for 2 h while allowing the temperature to warm to room temperature. The reaction was then quenched with saturated NH ₄ Cl and filtered through celite. The filtrate was extracted three times with DCM. The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE: EA = 1:1) to give 439B (360 mg) as a white solid: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.32 (d, J = 2.5 Hz, 1H), 7.64 (dd, J = 8.3, 2.5 Hz, 1H), 7.33 (d, J = 8.3 Hz, 1H), 3.65 (t, J = 9.4 Hz, 1H), 3.54 - 3.50 (m, 2H), 2.66 (ddd, J = 13.5, 9.4, 4.9 Hz, 1H), 2.32 - 2.15 (m, 1H). ESI (M+H) ⁺ = 197.2.

3-(6-chloropyridin-3-yl)-1-methylpyrrolidin-2-one (Example 439)

To a cold (0 °C) solution of 439B (210 mg, 1.1 mmol, 1.0 eq) in THF (10 mL) was added 60% NaH (64 mg, 1.6 mmol, 1.5 eq). The reaction mixture was stirred at 0° C. for 15 min, then iodomethane (0.053 mL, 0.8 mmol, 0.8 eq) in THF (0.5 mL) was added dropwise. The resulting solution was stirred at 0° C. for an additional 2 h and quenched with saturated NH ₄ Cl. The system was partitioned and separated and the aqueous phase was extracted three times with EA. The combined organic phases were _{dried over Na 2} SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 1:4) to give Example 439 (80 mg) as a brown oil. ESI (M+H) ⁺ = 211.2

The following examples were synthesized using the above procedure:

Scheme 75

1-(2-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)pyrimidin-5-yl)pyridin-2(1H) -On (Example 459)

From the starting material 2-chloro-5-iodopyrimidine, the title compound was obtained in a manner analogous to that described in General Method 5 and General Method 1 ^{: 1} H NMR (400 MHz, CD ₃ OD) δ 8.41-8.40 (d , J = 2.4 Hz, 2H), 8.34 (s, 2H), 7.63-7.59 (m, 2H), 6.63-6.61 (d, J = 8.8 Hz, 1H), 6.49-6.46 (m, 1H), 4.49- 4.41 (m, 2H), 2.40-2.39 (d, J = 1.2 Hz, 3H), 2.31-2.25 (m, 2H), 2.08-2.05 (t, J = 13.6 Hz, 2H), 1.69-1.64 (m, 2H).

The following examples were synthesized using the above procedure:

Scheme 76

(E)-3-(2-(methoxycarbonyl)phenyl)acrylic acid (461A)

A mixture of methyl 2-formylbenzoate (2.5 g, 15.23 mmol, 1.0 eq), malonic acid (1.8 g, 17.67 mmol, and 1.16 eq), morpholine (0.15 mL) and pyridine (4 mL) was heated at 100 °C. Stirred for 4 hours. After cooling to room temperature, the resulting solution was poured into a mixture of crushed ice (50 g) and 35% aqueous HCl (25 mL). The precipitate was filtered and washed with water (25 mLx2). The white solid was then recrystallized from methanol to give 461A (2.0 g, 63.7% yield): ¹ H NMR (400MHz, CDCl ₃ ) 8.58 (d, J = 15.9 Hz, 1H), 7.99 (dd, J = 7.8, 1.1 Hz, 1H), 7.61 - 7.68 (m, 1H), 7.53 - 7.60 (m, 1H), 7.41 - 7.51 (m, 1H), 6.33 (d, J = 15.9 Hz, 1H), 3.95 (s) , 3H).

(E)-methyl 2- (3-azido-3-oxoprop-1-en-1-yl) benzoate (461B)

To a solution of 461A (1.2 g, 5.82 mmol, 1.0 eq) and Et ₃ N (1.6 mL 11.64 mmol, 2.0 eq) in toluene (15 mL) was added DPPA (1.2 mL, 5.53 mmol, 0.95 eq) dropwise. The mixture was continued stirring at room temperature for 16 h. The solution was concentrated and purified by flash column (petroleum ether: EtOAc = 10:1) to give 461B as a white solid (1.0 g, 74.6% yield): ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56 (d, J = 15.8 Hz, 1H), 8.00 (dd, J = 7.8, 0.9 Hz, 1H), 7.53 - 7.67 (m, 2H), 7.42 - 7.53 (m, 1H), 6.31 (d, J = 15.8 Hz, 1H) ), 3.95 (s, 3H).

Methyl 1-oxo-1,2-dihydroisoquinoline-5-carboxylate (461)

A solution of 461B (500 mg, 2.16 mmol, 1.0 eq) in diphenylmethane (3 mL) was stirred under nitrogen at 80° C. for 1 h. After that, the mixture was continuously stirred at 240° C. for 3 hours. After cooling to room temperature, the mixture was purified by flash column (petroleum ether: EtOAc: 2: 1) to give the crude product, which was further purified by preparative HPLC to give Example 461 as a white solid (30 mg). , 6.8% yield): MS (ESI+) m/z 204.0 (M+H) ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.51 (br. s., 1H), 8.46 (d, J = 7.9 Hz, 1H), 8.27 (d, J = 6.9 Hz, 1H), 7.57 (t, J = 7.8 Hz, 1H), 7.24 - 7.46 (m, 2H), 3.90 (s, 3H).

Scheme 77

2-methoxy-5- (1-methyl-1H-pyrazol-4-yl) pyridine (462A)

(6-methoxypyridin-3-yl)boronic acid (820 mg, 5.4 mmol, 1.0 eq), 4-bromo-1-methyl-1H-pyrazole ( A suspension of 1.04 g, 6.4 mmol, 1.2 eq), Pd(dppf)Cl ₂ (392.3 mg, 0.54 mmol, 0.1 eq) and Cs ₂ CO ₃ (3.5 g, 10.8 mmol, 2.0 eq) _{under N 2} for 16 h Stirred at 110°C. The reaction mixture was partitioned between ethyl acetate and water. The organic phase was separated and washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 20:1) to give 462A (662.4 mg) as a yellow solid: ESI (M+H) ⁺ = 190.1.

5-(1-methyl-1H-pyrazol-4-yl)pyridin-2(1H)-one (Example 462)

To a solution of 462A (200 mg, 1.1 mmol) in EtOH (0.5 mL) was _{added HBr solution (40% in H 2} O, 2.5 mL). The reaction mixture was stirred at 80° C. for 20 h. The reaction mixture was cooled to room temperature and basified by dropwise addition of _{aqueous NH 3 .} The solvent was evaporated under reduced pressure and the crude residue was purified by flash chromatography (DCM: MeOH: NH ₄ OH) = 10:1:0.1) to give Example 462 (120 mg) as a gray solid. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.94 (dd, J = 9.4, 2.6 Hz, 1H), 7.77 (d, J = 2.3 Hz, 1H), 7.65 (s, 1H), 7.38 (d, J) = 1.1 Hz, 1H), 6.57 (dd, J = 21.7, 9.3 Hz, 1H), 3.76 (s, 3H). ESI (M+H) ⁺ = 176.1.

Scheme 78.

common method 13

( 1S, 3S )-N ^One -(5-iodopyridin-2-yl)-N ³ - (5- (methylthio) pyrimidin-2-yl) cyclopentane-1, 3-diamine (463A)

131C (150 mg, 0.669 mmol, 1.0 eq), 2-fluoro-5-iodopyridine (178.9 mg, 0.802 mmol, 1.2 eq) and K ₂ CO ₃ (277.2 mg, 2.006 mmol, 3.0 eq) was stirred at 140° C. under N ₂ for 16.5 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 1:1) to give 463A (125.5 mg). ESI (M+H) ⁺ = 428.1.

general method 14

3-methyl-1-(6-(((( 1S, 3S )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)imidazolidine-2,4-dione (463)

463A (65.5 mg, 0.153 mmol, 1.0 eq), 3-methylimidazolidine-2,4-dione (35.0 mg, 0.307 mmol, 2.0 eq), N ₁ , N _{2 in} i-PrOH (3 mL) A suspension of dimethylcyclohexane-1,2-diamine (10.9 mg, 0.077 mmol, 0.5 eq), CuI (14.6 mg, 0.077 mmol, 0.5 eq) and K ₃ PO ₄ (97.6 mg, 0.460 mmol, 3.0 eq) in N ₂ was purged. The reaction mixture was stirred at 110° C. under microwave irradiation for 4 hours. The reaction mixture was concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 20:1) to give Example 463 (20 mg) as a white powder: ¹ H NMR (400 MHz, TFA- d ₄ ) δ 8.8 ( brs, 2H), 8.40 (d, J = 2.4 Hz, 1H), 8.25 (dd, J = 9.8, 2.3 Hz, 1H), 7.23 (d, J = 9.8 Hz, 1H), 4.83-4.78 (m, 1H) ), 4.68 (s, 2H), 4.56-4.39 (m, 1H), 3.28 (s, 3H), 2.62 - 2.49 (m, 2H), 2.54 (s, 3H), 2.45 - 2.30 (m, 2H), 1.96-1.88 (m, 2H). ESI (M+H) ⁺ = 414.3.

The following examples were synthesized using the above procedure:

Scheme 79

( 1S, 3S )-N ^One -(5-(2-chlorophenyl)pyridin-2-yl)-N ³ -(5-(methylthio)pyrimidin-2-yl)cyclopentane-1,3-diamine (Example 467)

From the starting materials 463A and (2-chlorophenyl)boronic acid, the title compound was obtained in a manner analogous to that described in General Method 4 ^{: 1} H NMR (400 MHz, CDCl ₃ ) δ 10.20 (s, 1H), 8.53 (brs). , 1H), 8.45 (s, 2H), 7.95 (dd, J = 9.2, 1.4 Hz, 1H), 7.83 (s, 1H), 7.56 - 7.47 (m, 1H), 7.36 (dd, J = 5.9, 3.4 Hz, 2H), 7.25-7.27 (m, 1H), 7.13 (d, J = 9.3 Hz, 1H), 4.59 (brs, 1H), 4.24 (brs, 1H), 2.52 - 2.29 (m, 5H), 2.20 (ddt, J = 20.7, 13.8, 7.0 Hz, 2H), 1.99 - 1.86 (m, 1H), 1.74 (dt, J = 13.0, 6.6 Hz, 1H). ESI (M+H) ⁺ = 412.3.

Scheme 80

(One S , 3 S )-N ^One -(5-cyclopropylpyrimidin-2-yl)-N ³ -(5-iodopyridin-2-yl) cyclopentane-1, 3-diamine (468A)

From the starting material 425C (44.5 g, 208.7 mmol, 1.0 eq), 425D (24.7 g) was obtained as a pale solid in a similar manner to that described above. LCMS [M+H] ⁺ =422.

2-(6-(((1 S , 3 S )-3-((5-cyclopropylpyrimidin-2-yl) amino) cyclopentyl) amino) pyridin-3-yl) pyridazin-3(2H)-one (Example 468)

468A (18.7 g, 44.4 mmol, 1.0 eq), pyridazin-3(2H)-one (8.53 g, 88.8 mmol, 2.0 eq), (1 S , 2 S ) -N ¹ , N ^{2 in} DMSO (150 mL) - In a solution of dimethylcyclohexane-1,2-diamine (1.26 g, 8.88 mmol, 0.2 eq) and CuI (0.85 g, 4.44 mmol, 0.1 eq) K ₂ CO ₃ (18.5 g, 133.2 mmol, 3.0 eq) was added was added and the resulting system was stirred at 135° C. for 12 h under _{N 2 .} The reaction mixture was cooled to room temperature, poured into water (1 L) and extracted with ethyl acetate (150 mL x3). The combined organic layers were washed with brine and dried over anhydrous Na ₂ SO ₄ . The ethyl acetate phase was filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: eluting with MeOH = 100:1 to 50:1) to give Example 468 (25.3 g) as a pale yellow solid: ¹ H NMR (400 MHz, CD ₃ OD) ) δ 8.15 (d, J = 2.5 Hz, 1H), 8.07 (s, 2H), 8.02 (dd, J = 3.9, 1.6 Hz, 2H), 7.60 (dd, J = 9.0, 2.6 Hz, 1H), 7.46 (dd, J = 9.4, 3.9 Hz, 1H), 7.06 (dd, J = 9.5, 1.6 Hz, 1H), 6.59 (d, J = 9.0 Hz, 1H), 4.42 - 4.27 (m, 2H), 2.34 - 2.18 (m, 2H), 1.99 (t, J = 6.8 Hz, 2H), 1.81 - 1.70 (m, 1H), 1.65 - 1.53 (m, 2H), 0.95 - 0.85 (m, 2H), 0.66 - 0.54 ( m, 2H). LCMS [M+H] ⁺ = 390.

The following examples were synthesized using the above procedure:

Scheme 74

2-oxo-1,2-dihydroquinoline-5-carboxylic acid (478A)

A suspension of 2-chloroquinoline-5-carboxylic acid (300 mg, 1.45 mmol, 1.0 eq) in AcOH (5 mL) and H _{2 O (2 mL) was stirred at 130° C. overnight.} The reaction was cooled to 0° C. and stirred for 0.5 h. The precipitate was collected by filtration and the solid was dried in vacuo to give 478A (250 mg).

1-(6-(((1 S , 3 S )-3-((5-cyclopropylpyrimidin-2-yl) amino) cyclopentyl) amino) pyridin-3-yl)-2-oxo-1,2-dihydroquinoline-5-carboxylic acid (Example 478 )

478A (18 mg, 0.1 mmol, 1.0 eq), (1S,3S)-N1-(5-cyclopropylpyrimidin-2-yl)-N3-(5-iodopyridin-2-yl) in DMSO (3mL) ) cyclopentane-1,3-diamine (40 mg, 0.1 mmol, 1.0 eq), quinolin-8-ol (3 mg, 0.02 mmol, 0.2 eq), CuI (4 mg, 0.02 mmol, 0.2 eq), K ₂ A suspension of CO ₃ (20 mg, 0.15 mmol, 1.5 eq) was purged with _{N 2 .} The reaction mixture was then stirred at 120° C. for 2 hours under microwave irradiation. The reaction mixture was filtered and the filtrate was partitioned between ethyl acetate (5 mL) and water (10 mL). After separation, the aqueous phase was concentrated under reduced pressure. The crude residue was purified by preparative HPLC to give Example 478 (4.7 mg): ¹ H NMR (400 MHz, CD ₃ OD) δ 9.11 (d, J = 10.1 Hz, 1H), 8.25 (s, 2H) , 8.05 (s, 1H), 7.95 (d, J = 7.5 Hz, 1H), 7.79 (d, J = 9.4 Hz, 1H), 7.67 - 7.50 (m, 1H), 7.27-7.15 (m, 2H), 6.84 (d, J = 10.1 Hz, 1H), 4.58-4.45 (m, 1H), 4.38-4.28 (m, 1H), 2.53 - 2.28 (m, 2H), 2.27 - 2.06 (m, 2H), 1.91- 1.68 (m, 3H), 1.03-0.92 (m, 2H), 0.73-0.63 (d, J = 4.5 Hz, 2H). ESI (M+H) ⁺ = 483.

The following examples were synthesized using the above procedure:

Scheme 75

Common Method 15

6'-((( 1S, 3S )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-4-(2H-tetrazol-5-yl)-2H-[1, 3′-bipyridine ]-2-one (Example 481)

A mixture of 451 (35 mg, 0.083 mmol, 1.0 eq), NH ₄ Cl (44 mg, 0.83 mmol, 10.0 eq) and NaN ₃ (54 mg, 0.83 mmol, 10.0 eq) in DMF (2 mL) at 100° C. Stirred for 5 hours. The mixture was cooled to room temperature and filtered. The filtrate was purified by preparative HPLC to give Example 481 (13 mg, 33.7%) as a yellow solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm: 8.37 (s, 2H), 8.15 (d, J = 2.0 Hz, 1H), 7.98 (dd, J = 2.1 Hz, 9.6 Hz, 1H), 7.83 ( dd, J = 0.4 Hz, 7.2 Hz, 1H), 7.34 (d, J = 1.2 Hz, 1H), 7.10-7.15 (m, 2H), 4.46-4.50 (m, 1H), 4.27-4.31 (m, 1H) ), 2.43-2.43 (m, 5H), 2.11-2.18 (m, 2H), 1.71-1.80 (m, 2H). LCMS [M+H] ⁺ = 463.4.

The following examples were synthesized using the above procedure:

Scheme 76

Methyl 1-methyl-6-oxo-1, 6-dihydropyridine-3-carboxylate (484A)

A solution of methyl 2-oxo-2H-pyran-5-carboxylate (3 g, 19.465 mmol, 1.0 eq) and methylamine (33% in EtOH, 1.904 g, 20.439 mmol, 1.05 eq) in EtOH (10 mL) was stirred at 60° C. for 18 h in a sealed tube. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 1:2) to give 484A (393.6 mg) as a yellow solid. ESI (M+H) ⁺ = 168.1

Methyl 5-bromo-1-methyl-6-oxo-1, 6-dihydropyridine-3-carboxylate (484B)

A solution of 484A (393.6 mg, 2.355 mmol, 1 eq) and NBS (628.6 mg, 3.532 mmol, 1.5 eq) in AcOH (16 mL) was stirred at 80° C. for 2 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE: EA = 1:1) to give 484B (377.3 mg) as a white solid: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.26 (d, J = 2.3 Hz, 1H), 8.18 (d, J = 2.3 Hz, 1H), 3.90 (s, 3H), 3.68 (s, 3H). ESI (M+H) ⁺ = 246.0.

Methyl 6′-chloro-1-methyl-2-oxo-1,2-dihydro-[3,3′-bipyridine]-5-carboxylate (484C)

484C was obtained from the starting materials 484B and (6-chloropyridin-3-yl)boronic acid in a similar manner to that described above ^{: 1} H NMR (400 MHz, CDCl ₃ ) δ 8.68 (d, J = 2.2 Hz, 1H) , 8.27 (d, J = 2.3 Hz, 1H), 8.21 - 8.13 (m, 1H), 8.08 (d, J = 2.4 Hz, 1H), 7.39 (t, J = 11.7 Hz, 1H), 3.91 (s, 3H), 3.69 (s, 3H). ESI (M+H) ⁺ = 279.2

methyl 6'-(((1 S , 3 S )-3-((tert-butoxycarbonyl) amino) cyclopentyl) amino)-1-methyl-2-oxo-1,2-dihydro-[3, 3′-bipyridine]-5-carboxyl Rate (484D)

The starting material 484C and tert- butyl ((1S, 3S) -3- amino-cyclopentyl) 484D in a manner similar to that described above from the carbamate was obtained: ESI (M+H) = 443.3

methyl 6'-(((1 S , 3 S )-3-Aminocyclopentyl) amino)-1-methyl-2-oxo-1,2-dihydro-[3, 3′-bipyridine]-5-carboxylate (484E)

To a solution of 484D (35 mg) in DCM (1 mL) was added TFA (1 mL) dropwise. The reaction mixture was stirred at room temperature for 30 min and then concentrated under reduced pressure. The residue was dissolved in MeOH and then the pH level was adjusted to 8 by addition of an ^{ion exchange resin (Ambersep® 900 OH - form).} The mixture was filtered and the filtrate was concentrated under reduced pressure to give 484E (27 mg) 8 as a yellow oil.

methyl 1-methyl-6'-(((1 S , 3 S )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-1,2-dihydro-[3,3'-bipyridine]-5- Carboxylate (484F)

From the starting materials 484E and 2-chloro-5-(methylthio)pyrimidine, 484F was obtained in an analogous manner to that described above: ESI (M+H) ⁺ = 467.2.

1-methyl-6'-(((1 S , 3 S )-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-1,2-dihydro-[3,3'-bipyridine]-5- Carboxylic Acid (Example 484)

Example 484 was obtained from the starting material 484F in a similar manner as described above ^{: 1} H NMR (400 MHz, CD ₃ OD) δ 8.51 (dd, J = 14.0, 2.1 Hz, 2H), 8.35 (s, 2H), 8.25 (dd, J = 10.7, 2.2 Hz, 2H), 7.10 (d, J = 9.5 Hz, 1H), 4.51 - 4.41 (m, 1H), 4.31 - 4.20 (m, 1H), 3.69 (s, 3H), 2.49 - 2.39 (m, 1H), 2.35 (d, J = 7.6 Hz, 3H), 2.33 - 2.27 (m, 1H), 2.22 - 2.07 (m, 2H), 1.84 - 1.65 (m, 2H). ESI (M+H) ⁺ = 453.3.

Scheme 77

6'-(((1S,3S)-3-aminocyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (485)

485 was obtained from the starting material 6'-chloro-2H-[1,3'-bipyridin]-2-one in a manner analogous to that described in General Method 2 and General Procedure 6 : m/z 271.0 [M+H ] ⁺ .

The following examples were synthesized using the above procedure:

Scheme 78

6'-(((1 S , 3 S )-3-(thieno[3,2-d]pyrimidin-2-ylamino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (487)

485 (150 mg, 0.405 mmol, 1.0 eq), DIPEA (260 mg, 2.0 mmol, 5.0 eq) and 2-chlorothieno[3,2-d]pyrimidine (69 mg, 0.405 mmol) in DMA (2 mL) , 1.0 eq) was stirred at 150 °C for 30 min. The resulting mixture was diluted with ethyl acetate (10 mL), washed with water (2×10 mL) and the aqueous phase was extracted with ethyl acetate (4×20 mL). The combined organic layers were concentrated in vacuo and purified by preparative HPLC to give 487 (12 mg) as a white solid: LCMS m/z 405 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.73 (brs, 1H), 10.21 (brs, 1H), 9.03 (brs, 1H), 8.19 (d, J = 5.3 Hz, 1H), 7.92 (dd, J = 9.4, 2.3 Hz, 1H), 7.83 (d, J = 2.3 Hz, 1H), 7.54 - 7.43 (m, 2H), 7.41 (d, J = 5.5 Hz, 1H), 7.26 - 7.21 (m, 1H), 6.69 (dd, J = 9.3, 1.2 Hz, 1H), 6.38 - 6.29 (m, 1H), 4.81 - 4.59 (m, 1H), 4.53 - 4.25 (m, 1H), 2.52 - 2.31 (m, 3H), 2.08 (dt, J = 14.3, 7.8 Hz, 1H), 1.98 - 1.77 (m, 2H).

The following examples were synthesized using the above procedure:

Scheme 79

2-(6-(((1S, 3S)-3-(thieno[2,3-d]pyrimidin-2-ylamino)cyclopentyl)amino)pyridin-3-yl)pyridazine-3(2H )-on (489)

A solution of 486 (50 mg, 0.18 mmol), 5-chlorothiazolo[4,5-d]pyrimidine (30.5 mg, 0.18 mmol), DIPEA (70 mg, 0.54 mmol) in DMSO (5 mL) was added overnight with N ₂ under stirring at 100°C. The reaction mixture was extracted with EA and water. The organic phase was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by preparative TLC and preparative HPLC to give 2.6 mg of 489 (2.3 mg): ESI (M+H) ⁺ = 406.48; ¹ H NMR (400 MHz, CD ₃ OD) δ 8.76 (s, 1H), 8.39 (d, J = 2.2 Hz, 1H), 8.22 (d, J = 9.7 Hz, 1H), 8.10 (d, J = 3.8) , 1.6 Hz, 1H), 7.52 (dd, J = 9.5, 3.9 Hz, 1H), 7.22 (d, J = 6.0 Hz, 2H), 7.16 - 7.06 (m, 2H), 4.67 - 4.48 (m, 1H) , 4.36 - 4.23 (m, 1H), 2.52 - 2.31 (m, 2H), 2.30 - 2.12 (m, 2H), 1.86 - 1.69 (m, 2H).

The following examples were synthesized using the above procedure:

Scheme 80

2-chloro-7-methylthieno [3, 2-d] pyrimidine (494A)

2,4-dichloro-7-methylthieno[3,2-d]pyrimidine (500 mg, 2.3 mmol, 1 eq), Pd(OH) _{2 in EA (8 mL) and i-PrOH (1 mL)} A suspension of (20% on carbon, 200 mg, 0.14 mmol, 0.55 eq) and NaOAc (400 mg, 4.8 mmol, 2.0 eq _{) was stirred in a Parr apparatus under H 2} atmosphere (50 psi) at room temperature for 18 h. The reaction mixture was filtered over Celite and the filtrate was concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 4:1) to give 494A (320 mg) as a white powder. ESI (M+H) ⁺ = 185.0; ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.09 (s, 1H), 7.73 (q, J = 1.1 Hz, 1H), 2.51 (d, J = 1.2 Hz, 3H).

6'-(((1 S , 3 S )-3-((7-methylthieno[3,2-d]pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (494) )

A suspension of 494A (28.45 mg, 0.1541 mmol, 1.0 eq), 485 (50 mg, 0.185 mmol, 1.2 eq) and K ₂ CO ₃ (63.9 mg, 0.462 mmol, 3.0 eq) in DMSO (5 mL) under _{N 2} Stirred at 140° C. for 16 hours. The reaction mixture was extracted with DCM and water. The organic phase was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 20:1) to give 494 (10.4 mg) as a white powder. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.92 (d, J = 2.6 Hz, 1H), 7.60 (dd, J = 6.8, 1.8 Hz, 1H), 7.47 (ddd, J = 9.0, 6.6, 2.1 Hz, 1H), 7.39 (dd, J = 8.9, 2.7 Hz, 1H), 7.21 (d, J = 7.2 Hz, 1H), 6.92 (d, J = 7.0 Hz, 1H), 6.53 (d, J = 9.1) Hz, 1H), 6.44 (d, J = 8.9 Hz, 1H), 6.27 (td, J = 6.7, 1.3 Hz, 1H), 4.52 - 4.39 (m, 1H), 4.38 - 4.26 (m, 1H), 2.27 (d, J = 0.9 Hz, 3H), 2.16 (dd, J = 9.8, 5.4 Hz, 2H), 2.03 - 1.86 (m, 3H), 1.63 - 1.47 (m, 1H). ESI (M+H) ⁺ = 419.3

The following examples were synthesized using the above procedure:

Scheme 81

1- (2-chloropyrimidin-5-yl) ethan-1-ol (496)

To a solution of 1-(2-chloropyrimidin-5-yl)ethan-1-one (500 mg, 3.2 mmol, 1.0 eq) in MeOH (15 mL) was added NaBH ₄ (240 mg, 6.4 mmol, 2.0 eq) was added at 0°C. After stirring at room temperature for 2 h, water (5 mL) was added to the mixture to quench the reaction. The resulting mixture was concentrated under reduced pressure, and the crude residue was purified by silica gel column chromatography (hexane:ethyl acetate = 5:1 to ethyl acetate) to give 496 as a pale solid (100 mg). ESI [M+H] ⁺ = 159.1.

Scheme 82

2-(6-(((1S,3S)-3-((5-(methylthio)pyridin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)pyridazine-3(2H)- On (497)

For starting materials 416 and 486 , a procedure analogous to that described in General Method 2 gave 497 : ¹ H NMR (400 MHz, CD ₃ OD) δ 8.28 (d, J = 2.3 Hz, 1H), 8.04 (dd, J) = 9.6, 2.3 Hz, 1H), 7.97 (dd, J = 3.8, 1.5 Hz, 1H), 7.82 (dd, J = 9.4, 2.2 Hz, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.40 (dd, J = 9.5, 3.9 Hz, 1H), 7.00 (ddd, J = 9.6, 6.5, 5.0 Hz, 3H), 4.37 - 4.20 (m, 2H), 2.39 (s, 3H), 2.37 - 2.29 (m) , 2H), 2.17 (t, J = 6.4 Hz, 2H), 1.80 - 1.67 (m, 2H). ESI (M+H) ⁺ = 395.3

The following examples were synthesized using the above procedure:

Scheme 83

General Procedure 16

6'-(((1S, 3S)-3-((5-(methylthio) pyrimidin-2-yl) amino) cyclopentyl) amino)-[3, 3'-bipyridin]-4-ol ( Example 499)

To a solution of compound 402 (40 mg, 0.1 mmol, 1eq) in NMP (3 mL) was added LiCl (42 mg, 1 mmol, 10 eq) and p -toluenesulfonic acid (172 mg, 1 mmol, 10 eq) . The mixture was stirred at 180° C. for 4 h and cooled to room temperature. The mixture was diluted with water (10 mL) and then _{basified with saturated NaHCO 3} to pH=10. The resulting mixture was extracted with ethyl acetate (10 mL x 6), and the combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The resulting residue was purified by preparative HPLC to give Example 499 (10 mg, 26%) as a yellow solid: LCMS [M+H] ⁺ = 395; ¹ H NMR (400 MHz, CD ₃ OD) δ : 8.33 (s, 2H), 8.18 (s, 1H), 7.88 (s, 1H), 7.77-7.72 (m, 1H), 6.63 (d, J = 9.2) Hz, 1H), 6.52 (d, J = 9.2 Hz, 1H), 4.45-4.35 (m, 2H), 2.35 (s, 3H), 2.30-2.24 (m, 2H), 2.04-1.98 (m, 2H) , 1.63-1.60 (m, 2H).

The following examples were synthesized using the above procedure:

Scheme 84

(One S , 3 S )-N ^One -(5-(difluoromethoxy)pyrimidin-2-yl)-N3-(5-nitropyridin-2-yl)cyclopentane-1,3-diamine (500A)

411 (43 g, 176 mmol, 1.0 eq), 2-chloro-5-nitropyridine (27.9 g, 176 mmol, 1.0 eq) and K ₂ CO ₃ (48.6 g, 382 mmol, 2.0 eq) in DMSO (500 mL) ) was stirred at 80 °C for 16 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was partitioned between EA (400 mL) and water (400 mL); The aqueous phase was extracted with EA (300 mL). The combined organic phases were washed with brine (400 mL) and then concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (PE:EA=3:1) to give 500A (52 g).

N ² -((1S, 3S)-3-((5-(difluoromethoxy) pyrimidin-2-yl) amino) cyclopentyl) pyridine-2,5-diamine (500B)

To a solution of 500A (45 g, 123 mmol, 1.0 eq) in MeOH (450 mL) was added 10% Pd/C (4.5 g). The reaction mixture was degassed 3 times with H ₂ and stirred for 8 hours at room temperature under H _2. The reaction mixture was filtered through Celite. The filtrate was concentrated to remove the solvent, and the residue was purified by column chromatography on silica gel (EA:MeOH=30:1) to give 500B (31 g).

3-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-1-methyl is Midazolidine-2,4-dione (Example 500)

A solution of 500B (27.2 g, 81 mmol, 1.0 eq) and 4-nitrophenyl carbonochloridate (16.3 g, 81 mmol, 1.0 eq) in acetonitrile (280 mL) was stirred at room temperature for 1 h. Methyl methylglycinate hydrochloride (11.8 g, 84 mmol, 1.1 eq) and DIPEA (31.3 g, 24.2 mmol, 3.0 eq) were added to the reaction and stirred for an additional 16 h. The reaction mixture was concentrated to remove most of the solvent and the residue was partitioned between DCM (70 mL) and water (70 mL). The organic phase was separated and concentrated. The resulting residue was purified by column chromatography on silica gel (DCM: MeOH=40:1) to give the crude product as a slurry. The compound was further purified by trituration in PE/EA (4:1, 40 mL) and decolorized with activated charcoal in MeOH to give Example 500 (25 g): ¹ H NMR (400 MHz, CD ₃ OD) ) δ 8.16 (s, 2H), 7.91 (d, J = 2.1 Hz, 1H), 7.36 (dd, J = 9.0, 2.6 Hz, 1H), 6.93 - 6.43 (m, 2H), 4.47 - 4.25 (m, 2H), 4.08 (s, 2H), 3.02 (s, 3H), 2.35 - 2.19 (m, 2H), 2.12 - 1.87 (m, 2H), 1.69 - 1.46 (m, 2H).

Scheme 85

2-chloro-6-methylthieno [3, 2-d] pyrimidine (501A)

2,4-dichloro-6-methylthieno[3,2-d]pyrimidine (1.5 g, 6.85 mmol, 1.0 eq), zinc (1.8 g, 27.39 mmol, 4.0 eq) and acetic acid in methanol (30 mL) (2.4 mL, 41.08 mmol, 6.0 eq) was stirred at 70° C. for 16 h. After cooling to room temperature, the mixture was filtered and the filter cake was washed with methanol (30 mL×2). The filtrate was concentrated and purified by flash column (petroleum ether: EtOAc = 4:1) to give 501A (620 mg, 47.6% yield) as a white solid. MS (ESI+) m/z 185.0 (M+H) ⁺

6- (bromomethyl) -2-chlorothieno [3,2-d] pyrimidine (501B)

A solution of 501A (600 mg, 3.25 mmol, 1.0 eq), N-bromosuccinimide (694.0 mg, 3.90 mmol, 1.2 eq) and AIBN (26.7 mg, 0.16 mmol, 0.05 eq) in carbon tetrachloride (20 mL) was It was refluxed in a sealed tube for 16 hours. After cooling to room temperature, the mixture was concentrated and purified by flash column (petroleum ether: EtOAc: 4:1) to give 501B (614 mg, 71.2% yield) as a yellow solid. MS (ESI+) m/z 262.9 (M+H) ⁺

(2-chlorothieno [3,2-d] pyrimidin-6-yl) methyl acetate (501C)

A solution of 510B (600 mg, 2.3 mmol, 1.0 eq) and cesium acetate (2.2 g, 11.4 mmol, 5.0 eq) in DMF (15 mL) was stirred at room temperature for 30 min. The mixture was diluted with EtOAc (100 mL), washed with water (80 mL) and brine (60 mL), and dried over Na ₂ SO ₄ . After filtration, the filtrate was concentrated and purified by flash column (petroleum ether: EtOA: 4:1) to give 501C (373 mg, 67.5% yield) as a white solid. MS (ESI+) m/z 243.0 (M+H) ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.07 (s, 1H), 7.39 - 7.52 (m, 1H), 5.43 (d, J = 0.9 Hz, 2H), 2.17 (s, 3H).

(2-chlorothieno [3, 2-d] pyrimidin-6-yl) methanol (501D)

A solution of 501C (160 mg, 0.66 mmol, 1.0 eq) and potassium carbonate (136.7 mg, 0.99 mmol, 1.5 eq) in methanol (15 mL) was stirred at room temperature for 1 h. Then the mixture was concentrated and purified by flash column (petroleum ether: EtOAc: 1:1) to give 501D (100 mg, 75.8% yield) as a white solid: MS (ESI+) m/z 201.0 (M+H) ) ⁺

6-(((tert-butyldimethylsilyl)oxy)methyl)-2-chlorothieno[3,2-d]pyrimidine (501E)

A solution of 501D (100 mg, 0.50 mmol, 1.0 eq) in DMF (5 mL) was cooled to 0 °C. TBSCl (97.6 mg, 0.65 mmol, 1.3 eq) was added followed by triethylamine (0.14 mL, 1.00 mmol, 2.0 eq). The mixture was stirred at room temperature for 16 h. The mixture was diluted with ethyl acetate (60 mL), washed with water (60 mL) and brine (30 mL), and dried over Na ₂ SO ₄ . The filtrate was concentrated and purified by flash column (petroleum ether: EtOAc: 4:1) to give 501D (126 mg, 80.5% yield) as a white solid: MS (ESI+) m/z 315.2 (M+H) ⁺

6'-(((1S,3S)-3-((6-(((tert-butyldimethylsilyl)oxy)methyl)thieno[3,2-d]pyrimidin-2-yl)amino)cyclopentyl )Amino)-2H-[1,3'-bipyridin]-2-one (501F)

In starting materials 501E and 485 , 501F was obtained in a manner analogous to that described in general method 5:

6'-(((1S, 3S)-3-((6-(hydroxymethyl) thieno[3,2-d]pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1 ,3'-bipyridin]-2-one (501)

TBAF (3 mL) was added to a solution of 501F (crude, 0.16 mmol, 1.0 eq) in DMSO (10 mL) and stirred for 30 min. The mixture was then diluted with water (50 mL), extracted with EtOAc (50 mL× ₂ ) and dried over Na 2 SO ₄ . The filtrate was concentrated and purified by flash column on silica gel (DCM: MeOH: 15:1) to give the crude compound, which was further purified by preparative HPLC to give pure 501 (2.3 mg, 3.3% yield by 2 steps) as white Obtained as a solid: MS (ESI+) m/z 435.1 (M+H)+

¹ H NMR (400 MHz, CD ₃ OD) δ 8.84 (s, 1H), 7.98 (d, J = 2.0 Hz, 1H), 7.82 (dd, J = 9.6, 2.4 Hz, 1H), 7.47 - 7.61 (m, 2H), 7.06 (d, J = 0.8 Hz, 1H), 6.99 (d, J = 9.5 Hz, 1H), 6.55 (dt, J = 9.1, 1.0 Hz, 1H), 6.41 (td, J = 6.8, 1.3) Hz, 1H), 4.83 (d, J = 1.1 Hz, 2H), 4.50 (t, J = 6.8 Hz, 1H), 4.17 - 4.28 (m, 1H), 2.22 - 2.39 (m, 2H), 2.07 - 2.17 (m, 2H), 1.63 - 1.77 (m, 2H).

Scheme 86

Tert-Butyl ((1 S , 3 S )-3-((5-bromopyridin-2-yl)oxy) cyclopentyl) carbamate (502A)

tert-Butyl ((1 S , 3 R )-3-hydroxycyclopentyl) carbamate (865 mg, 4.3 mmol, 1.5 eq) and 5-bromopyridin-2-ol (500 mg) in THF (20 mL) , 2.9 mmol, 1.0 eq) was added PPh ₃ (1.884 g, 7.2 mmol, 2.5 eq) to a cold (0° C.) solution, and then DEAD (1.04 mL, 7.2 mmol, 2.5 eq) under _{N 2 was added dropwise.} The reaction mixture was stirred at 0° C. for 1.5 h. The reaction mixture was then partitioned between EA and water. The separated organic phase was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 5:1) to give 502A (1.02 g) as a white solid: ESI (M+H) ⁺ = 357.2. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.15 (d, J = 2.4 Hz, 1H), 7.60 (ddd, J = 13.9, 7.8, 6.1 Hz, 1H), 6.58 (d, J = 8.8 Hz, 1H) , 4.50 (s, 1H), 4.20 (s, 1H), 2.20 (tt, J = 12.3, 6.3 Hz, 3H), 1.87 - 1.72 (m, 2H), 1.58 (d, J = 13.9 Hz, 1H), 1.44 (s, 9H).

(1S, 3S)-3-((5-bromopyridin-2-yl)oxy)cyclopentan-1-amine (502B)

From the starting material 502A , 502B was obtained as a yellow oil in a manner analogous to that described in General Method 6.

N-((1S, 3S)-3-((5-bromopyridin-2-yl)oxy)cyclopentyl)-5-(methylthio)pyrimidin-2-amine (502C)

From the starting material 502B and 2-chloro-5-(methylthio)pyrimidine 502C was obtained as a yellow solid in ^{a manner analogous to that described in General Method 5: ESI (M+H) +} = 381.1

6′-(((1S, 3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)oxy)-2H-[1, 3′-bipyridine]-2- On (502)

From the starting material 502 and pyridin-2(1H)-one , in a manner analogous to that described in general method 14 , 502 was obtained as a white solid: ESI (M+H) ⁺ = 396.3. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.40 (s, 2H), 8.21 - 8.14 (m, 1H), 7.74 (dd, J = 8.8, 2.8 Hz, 1H), 7.66 - 7.57 (m, 2H) , 6.90 (dd, J = 8.8, 0.5 Hz, 1H), 6.63 (dt, J = 8.9, 1.1 Hz, 1H), 6.49 (td, J = 6.8, 1.3 Hz, 1H), 5.61 - 5.53 (m, 1H) ), 4.51 (p, J = 7.1 Hz, 1H), 2.39 (s, 3H), 2.37 - 2.26 (m, 3H), 2.08 - 1.99 (m, 1H), 1.91 (tdd, J = 10.0, 7.7, 2.7 Hz, 1H), 1.74 - 1.60 (m, 1H).

Scheme 87

tert-butyl ((1 R ,4 R )-4-azidocyclopent-2-en-1-yl)carbamate (503A)

To a solution of tert-butyl ((1R,4S)-4-hydroxycyclopent-2-en-1-yl)carbamate (200 mg, 1.0 mmol, 1.0 eq) in THF (10 mL) DPPA (414 mg , 1.5 mmol, 1.5 eq) and DBU (230 mg, 1.5 mmol, 1.5 eq) were added at room temperature, followed by stirring for 2 days. Water (10 mL) was added to the solution, which was then extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered, concentrated and purified by chromatography on silica gel eluting with PE:EA=5:1 503A (200 mg, 89%) ) as a white solid.

Tert-Butyl ((1 R , 4 R )-4-Aminocyclopent-2-en-1-yl) carbamate (503B)

To a solution of 503A (200 mg, 0.9 mmol, 1.0 eq) in _{THF: H 2} O=4:1 (10 mL) was _{added PPh 3} (234 mg, 0.9 mmol, 1.0 eq) _{under N 2} atmosphere at room temperature. The mixture was then stirred at 50° C. overnight and then cooled to room temperature. The filtrate was concentrated and purified by chromatography on silica gel eluting with DCM: MeOH = 20:1 to 3:1 to give 503B (100 mg, 57%) as a semi-solid.

tert-Butyl ((1R,4R)-4-((5-(methylthio)pyrimidin-2-yl)amino)cyclopent-2-en-1-yl)carbamate (503C)

From the starting material 503B and 2-chloro-5-(methylthio)pyrimidine 503C was obtained as a brown solid in a manner analogous to that described in General Method 5.

(1R, 3R)-N1-(5-(methylthio)pyrimidin-2-yl)cyclopent-4-ene-1,3-diamine (503D)

503D was obtained as a brown oil from the starting material 503C in a manner analogous to that described in General Method 6 , which was used directly in the next step.

6′-(((1R, 4R)-4-((5-(methylthio) pyrimidin-2-yl) amino) cyclopent-2-en-1-yl) amino)-2H-[1,3 '-bipyridin]-2-one (503)

503 was obtained from the starting material 503D as an off-white solid in a manner analogous to that described in General Method 2 : LCMS [M+H] ⁺ = 393; ¹ H NMR (400 MHz, CD ₃ OD) δ 8.34 (s, 2H), 7.96 (s, 1H), 7.65-7.55 (m, 2H), 7.48-7.45 (m, 1H), 6.63-6.60 (m, 2H), 6.50-6.46 (m, 1H), 6.02-6.00 (m, 2H), 5.20-5.15 (m, 2H), 2.35 (s, 3H), 2.20-2.16 (m, 2H).

Scheme 88

Benzyl tert-butyl ((1R, 3R)-cyclopent-4-ene-1, 3-diyl) dicarbamate (504A)

In a solution of 503B (0.5 g, 2.5 mmol, 1.0 eq) in THF (10 mL) and H ₂ O (2 mL) _{Na 2} CO ₃ (0.65 g, 5.8 mmol, 2.5 eq) and Cbz-Cl (510 mg, 3.0 mmol, 1.2 eq) was added. The resulting solution was stirred at room temperature for 2.5 hours. The reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (25 mL x3). The combined organic layers were washed with brine and dried over anhydrous Na ₂ SO ₄ . The filtrate was concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (hexane:ethyl acetate = 5:1) to give 504A (566 mg) as a pale solid. ESI [M+H] ⁺ = 333.

Benzyl tert-butyl ((1R,3R)-4-hydroxycyclopentane-1,3-diyl)dicarbamate (504B-1) & benzyl tert-butyl ((1R,3R)-4-hydroxycyclopentane -1,3-diyl) dicarbamate (504B-2)

To a solution of 504A (524 mg, 1.7 mmol, 1 eq) in THF (10 mL) was _{added BH 3} .THF (1 N, 7.8 mL, 7.8 mmol, 4.5 eq) at 0 °C. The resulting solution was stirred at room temperature for 6 hours. H ₂ O (7.5 mL) and NaOH (3 M, 12 mL) were added followed by H ₂ O ₂ (30%, 20 mL). The mixture was stirred at room temperature for an additional 10 min, then EtOH (7.5 mL) was added. The resulting mixture was stirred at room temperature for 20 hours. After pouring into water (20 mL), the mixture was extracted with ethyl acetate (15 mL x 6). The combined organic layers were washed with brine and dried over anhydrous Na ₂ SO ₄ . The organic phase was filtered and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (hexane:ethyl acetate = 10:1 to 1:2) to give the title compound (566 mg in total) as a pale solid: ESI [M+H] ⁺ = 351.

tert-Butyl ((1R,3R)-3-amino-4-hydroxycyclopentyl)carbamate (504-1) & tert-butyl ((1R,4R)-4-amino-2-hydroxycyclopentyl) Carbamate (504-2)

360 mg of starting material (1.1 mmol, 1 eq) in MeOH (25 mL) was stirred with Pd(OH) ₂ (100 mg) _{under H 2 at room temperature for 19 h.} The catalyst was removed by filtration and the filtrate was concentrated to give the title compound as a pale solid. Yield: 200 mg. ESI [M+H] ⁺ = 217.

The following examples were synthesized using the procedure described above:

Scheme 89

tert-butyl ((1R,4S)-4-((tert-butyldiphenylsilyl)oxy)cyclopent-2-en-1-yl)carbamate (508A)

tert-Butyl ((1R,4S)-4-hydroxycyclopent-2-en-1-yl)carbamate (500 mg, 2.5 mmol, 1.0 eq) and imidazole (342 mg, To a cold (0°C) solution of 5.0 mmol, 2.0 eq) was added dropwise TBDPSCl (0.85 mL, 3 mmol, 1.3 eq). The reaction mixture was then stirred for 16 hours while the temperature was raised to room temperature. The reaction mixture was partitioned between EA and water. The separated organic layer was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by reverse phase flash chromatography (CH ₃ CN:H ₂ O = 40% to 90%) to give 508A (869.2 mg) as a clear oil: ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.65 (dd, J = 6.8, 1.0 Hz, 4H), 7.53 - 7.41 (m, 6H), 7.10 (d, J = 7.8 Hz, 1H), 4.67 (t, J = 6.7 Hz, 1H), 4.25 ( dd, J = 14.6, 7.2 Hz, 1H), 3.20 (d, J = 4.0 Hz, 1H), 2.53 (dt, J = 3.6, 1.8 Hz, 1H), 2.52 - 2.43 (m, 1H), 1.60 - 1.49 (m, 1H), 1.39 (s, 9H), 1.03 (s, 9H).

tert-butyl ((1R,2R,4S,5S)-4-((tert-butyldiphenylsilyl)oxy)bicyclo[3.1.0]hexan-2-yl)carbamate (508B)

_{To a cold (-15 °C) solution of Et 2} Zn (4.6 mL, 4.6 mmol, 3.0 eq) in dry DCM (5 mL) was added diiodomethane (0.74 mL, 9.2 mmol, 6 eq) in DCM (4 mL). was added dropwise. The reaction mixture was stirred at -15°C for 10 minutes until a white precipitate formed. 508A (670 mg, 1.5 mmol, 1.0 eq) in DCM (5 mL) was then added dropwise to the reaction mixture. Stirring was continued for 22 hours while the temperature was gradually warmed to room temperature. The reaction was quenched by addition of saturated NH ₄ Cl, which was then partitioned between DCM and water. The separated organic phase was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (PE:EA = 10:1) to give 508B (280 mg) as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.75 - 7.59 (m, 4H), 7.47 - 7.30 (m, 6H), 4.66 - 4.50 (m, 1H), 4.39 (td, J = 8.0, 4.8 Hz, 1H ), 4.01 (s, 1H), 2.17 - 2.07 (m, 1H), 1.44 (d, J = 7.8 Hz, 9H), 1.34 - 1.24 (m, 2H), 1.05 (d, J = 5.7 Hz, 9H) , 0.83 (dt, J = 9.0, 2.9 Hz, 1H), 0.46 - 0.33 (m, 1H).

(1R, 2R, 4S, 5S)-4-((tert-butyldiphenylsilyl)oxy)bicyclo[3.1.0]hexan-2-amine (508C)

To a solution of 508B (230 mg) in DCM (3 mL) was added TFA (5 mL). The reaction mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure. The residue was redissolved in MeOH and the pH level was increased to 8 by addition of ion exchange resin. The filtrate was concentrated under reduced pressure to give 508C (189.2 mg) as a yellow oil.

N-((1R,2R,4S,5S)-4-((tert-butyldiphenylsilyl)oxy)bicyclo[3.1.0]hexan-2-yl)-5-(methylthio)pyrimidine-2 -amine (508D)

504D was obtained as a brown oil from the starting material 504C and 2-chloro-5-(methylthio)pyrimidine in a manner analogous to that described in General Method 5.

(1S,2S,4R,5R)-4-((5-(methylthio)pyrimidin-2-yl)amino)bicyclo[3.1.0]hexan-2-ol (508E)

To a solution of 508D (50 mg) in THF (2 mL) was added pyridine HF (0.5 mL) dropwise. The reaction mixture was stirred at room temperature for 4 h, then partitioned between EA and water. The separated organic phase was washed with brine. The organic phase was _{dried over Na 2} SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel (DCM: MeOH = 10:1) to give 508E (54.4 mg) as a light yellow oil. ESI (M+H) ⁺ = 238.3

2-((1S,2R,4R,5R)-4-((5-(methylthio)pyrimidin-2-yl)amino)bicyclo[3.1.0]hexan-2-yl)isoindoline-1 ,3-dione (508F)

508E (50 mg, 0.2 mmol, 1.0 eq), isoindoline-1,3-dione (37 mg, 0.25 mmol, 1.2 eq) and PPh ₃ (600 mg, 2.3 mmol, 10.0 eq) in THF (10 mL) To a solution of DEAD (398 mg, 2.3 mmol, 10.0 eq) was added dropwise at 0° C. under _{N 2 .} The temperature was gradually raised to room temperature while the reaction mixture was stirred for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude residue was purified twice by flash chromatography on silica gel (PE:EA = 1:1) to give 508F (214 mg, contaminated with impurities) as a yellow oil. ESI (M+H) ⁺ = 367.4

(1R,2R,4R,5S)-N2-(5-(methylthio)pyrimidin-2-yl)bicyclo[3.1.0]hexane-2,4-diamine (508G)

A solution of 508F (214 mg, 0.584 mmol) and hydrazine hydrate (0.2 mL) in EtOH (5 mL) _{was stirred under N 2} at 60° C. for 2 h. The reaction mixture was concentrated under reduced pressure and purified by flash chromatography on silica gel (DCM:MeOH = 20:1) to give 508G (30 mg) as a yellow oil: ESI (M+H) ⁺ = 237.4

6′-(((1S,2R,4R,5R)-4-((5-(methylthio)pyrimidin-2-yl)amino)bicyclo[3.1.0]hexan-2-yl)amino)- 2H-[1,3'-bipyridin]-2-one (508)

From the starting material 508G , in a manner analogous to that described in General Method 2 , 508 was obtained as a yellow solid: ESI (M+H) ⁺ = 407.4. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.35 (s, 2H), 8.07 (d, J = 2.2 Hz, 1H), 7.93 (dd, J = 9.6, 2.4 Hz, 1H), 7.72 - 7.57 (m) , 2H), 7.12 (d, J = 9.5 Hz, 1H), 6.65 (d, J = 9.0 Hz, 1H), 6.51 (td, J = 6.8, 1.2 Hz, 1H), 4.23 (d, J = 5.9 Hz) , 1H), 2.36 (s, 3H), 2.22 - 2.10 (m, 1H), 1.93 (dt, J = 9.7, 4.3 Hz, 1H), 1.61 (dt, J = 9.9, 6.4 Hz, 2H), 0.76 ( dt, J = 7.8, 3.9 Hz, 1H), 0.69 (dd, J = 14.0, 7.9 Hz, 1H).

C. Biological Assays

Human recombinant PCSK9 was expressed as follows:

Protein sequence:

SEQ ID NO: 1

QEDEDGDYEELVLALRSEEDGLAEAPEHGTTATFHRCAKDPWRLPGTYVVVLKEETHLSQSERTARRLQAQAARRGYLTKILHVFHGLLPGFLVKMSGDLLELALKLPHVDYIEEDSSVFAQSIPWNLERITPPRYRADEYQPPDGGSLVEVYLLDTSIQSDHREIEGRVMVTDFENVPEEDGTRFHRQASKCDSHGTHLAGVVSGRDAGVAKGASMRSLRVLNCQGKGTVSGTLIGLEFIRKSQLVQPVGPLVVLLPLAGGYSRVLNAACQRLARAGVVLVTAAGNFRDDACLYSPASAPEVITVGATNAQDQPVTLGTLGTNFGRCVDLFAPGEDIIGASSDCSTCFVSQSGTSQAAAHVAGIAAMMLSAEPELTLAELRQRLIHFSAKDVINEAWFPEDQRVLTPNLVAALPPSTHGAGWQLFCRTVWSAHSGPTRMATAVARCAPDEELLSCSSFSRSGKRRGERMEAQGGKLVCRAHNAFGGEGVYAIARCCLLPQANCSVHTAPPAEASMGTRVHCHQQGHVLTGCSSHWEVEDLGTHKPPVLRPRGQPNQCVGHREASIHASCCHAPGLECKVKEHGIPAPQEQVTVACEEGWTLTGCSALPGTSHVLGAYAVDNTCVVRSRDVSTTGSTSEGAVTAVAICCRSRHLAQASQELQSGSGGLNDIFEAQKIEWHENLYFQGHHHHHH

The following analytical methods were used to identify and evaluate compounds of formula (I) that are effective in inhibiting PCSK9 function.

Example: PCSK9 SPR analysis

Surface plasmon resonance data ^{were collected on a Biacore™} T200 or 3000 system (GE Healthcare) at 25°C. Immobilization of streptavidin with HBS-N (10 mM HEPES, 0.15 M NaCl, pH 7.4) as running buffer at 25 °C using standard amine coupling chemistry to CM5 (GE Healthcare) or CMD500d sensor chip (XanTec Bioanalytics) did it Briefly, the carboxymethyl dextran surface was coated with 0.4 M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)/0.1 M N -hydride in a 1:1 ratio at a flow rate of 10 μL/min. It was activated by a 12 min infusion of roxy succinimide (NHS). For the capture of streptavidin, the protein was diluted to 0.2 mg/mL in 10 mM sodium acetate (pH 4.5) and captured by injecting 100 μL onto the activated chip surface. Residual activating groups were blocked by a 7 min injection of 1 M ethanolamine, pH 8.5. Avi-tagged PCSK9 protein was captured on the streptavidin surface by injecting 150 μL of protein diluted to 16 μg/mL in HBS-N, 0.05% tween-20, and 0.1 mM CaCl _{2 .} Typical surface densities obtained were 8000-10000 RU. SPR binding data were obtained using the appropriate dilution series of each compound at a flow rate of 30 μL/min, applying a capture time of 100 s and a dissociation time of 300 s. Running buffers for compound binding studies were HBS-N, 0.05% Tween-20, 0.1 mM CaCl ₂ , 4% DMSO. Data were corrected for volume effects without DMSO. All data were double referenced against blank injection and reference surfaces using standard processing procedures, and data processing and kinetics fitting were performed using Scrubber software, version 2.0c (BioLogic Software). Data were fitted using a simple 1:1 binding model to determine k _a , k _d and K _{D values.}

The ability of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to bind and inhibit PCSK9 was established with representative compounds of formula (I) listed in the table below:

Example: In vitro cellular assays to determine the effect of compounds on secreted PCSK9 levels, cellular LDLR levels and cell viability.

Compound screening was performed in 96-well tissue culture plates with 25,000 HepG2 cells plated in 200 μl of assay medium (DMEM (Gibco 31966-021) containing 10% lipoprotein-deficient FBS (Sigma S5394)). Cell plates were incubated overnight (20-24 hours), then assay medium was removed, cells were washed with 200 μl of DMEM, and 200 μl of compound or vehicle (0.3% DMSO) in assay medium was added to each well. . After 48 hours of incubation with the compounds, the following assays were performed.

For the measurement of secreted PCSK9 levels, a sample of 100 μl of cell culture assay medium was collected and stored at -80°C prior to analysis using the PCSK9 (human) AlphaLISA detection kit (Perkin Elmer AL270F). A sample (5 μl) of cell culture assay medium was transferred to a 384 well white optiplate (Perkin Elmer-6007290). Also included were 5 μl samples of assay medium alone to determine assay background and samples of assay medium supplemented with known concentrations of recombinant human PCSK9 (standard curve). 5 μl each of a solution of 20 μl of AlphaLISA anti-PCSK9 acceptor beads (final concentration 10 μg/mL) and biotinylated antibody anti-PCSK9 (final concentration 1 nM) diluted in AlphaLISA Immunoassay buffer (all provided in the AlphaLISA detection kit) was added to the sample of , and the plate was incubated at room temperature for 1 hour. After incubation, 25 μl of a solution of streptavidin donor beads (final concentration 40 μg/mL) diluted in AlphaLISA immunoassay buffer was added to each sample, and the samples were incubated for an additional 1 hour at room temperature protected from light. In the presence of PCSK9 (analyte), the donor and acceptor beads come into proximity. The emission at 615 nm is then measured in an Enspire Alpha plate reader after excitation. After determining the concentration of PCSK9 using the standard curve values, the percent inhibition was calculated using the following formula: (1-(test well value - mean background value)/(mean vehicle value - mean background value))*100

Cell viability assays were performed on the same cell plates as PCSK9 assays after cell media samples were collected. The assay is based on the reduction of MTS tetrazolium compounds by viable cells to produce a colored formazan product that is soluble in the cell culture medium. 20 μl of MTS reagent (Promega G543) was added to each cell well containing 100 μl of the remaining culture medium. In addition, wells containing cell-free MTS reagent and 100 μl of assay medium were included to determine background measurements. The plate was incubated at 37° C. for 1 hour and the optical density (OD) was measured at a wavelength of 490 nm. OD values were converted to % change in viability values using the formula: - (1-(test well values - mean background values)/(mean vehicle values - mean background values))*100

Cellular LDLR levels were determined using human LDL R immunoassays (R&D systems DLDLR0) and all reagents provided for immunoassays unless otherwise specified. HepG2 cells were treated as described above, and after 48 h of compound incubation, the medium was removed, cells were washed with phosphate buffered saline, and cells were washed with 50 µL of lysis buffer with protease inhibitor (Halt inhibitor cocktail - Pierce 78430). (25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40 and 5% glycerol). Lysates were removed by centrifugation and samples were stored at -80°C prior to analysis in immunoassays. In addition to the samples, a standard curve of known concentrations of recombinant LDLR diluted in calibration diluent was included and LDLR was not included to determine assay background. Samples (30 μl) were incubated in microplate well strips (pre-coated with capture antibody) with 50 μl of assay diluent for 2 h at room temperature. Then, the microplate well strips were washed 4 times with wash buffer, and 200 μl of human LDLR conjugate was added. After an additional 2 h incubation at room temperature, the plates were washed as before and 200 μl of the substrate solution was added. After 20 min, 50 μl of stop solution was applied, and the optical density of each well measured at 450 nm and a wavelength correction of 540 nm were applied. After determining the concentration of LDLR using the standard curve values, the percent increase was calculated using the following formula: (Test Well Value - Mean Background Value)/(Mean Vehicle Value - Mean Background Value)*100

As a positive control, PCSK9 translation inhibitor, R-4- (3 H - [ 1,2,3] triazolo [4,5- b] pyridin-3-yl) - N - (3- chloropyridin-2-yl ) - N - a (piperidin-3-yl) benzamide (SI-1) was synthesized according to WO 2014170786.

Using the above assays, the ability of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to inhibit PCSK9 function was established with representative compounds of formula (I) listed in the table below:

included for reference

All publications and patents mentioned herein are incorporated herein by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

equivalent

While specific embodiments of the invention have been discussed, the above specification is illustrative and not restrictive. Numerous modifications of the present invention will become apparent to those skilled in the art upon review of this specification and the claims that follow. The full scope of the invention should be determined by reference to this specification, along with such modifications, and by reference to the claims, along with equivalents of the full scope.

SEQUENCE LISTING <110> SERRANO-WU, MICHAEL H. CHAMBERS, MARK GOLDSMITH, ERICA TIERNEY, JASON JANDU, KARAMJIT CLARK, DAVID HINCHLIFFE, PAUL <120> PCSK9 INHIBITORS AND METHODS OF USE THEREOF <130> DGH-00101 <140> 16/744,929 <141> 2020-01-16 <150> 62/794,234 <151> 2019-01-18 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 693 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 1 Gln Glu Asp Glu Asp Gly Asp Tyr Glu Glu Leu Val Leu Ala Leu Arg 1 5 10 15 Ser Glu Glu Asp Gly Leu Ala Glu Ala Pro Glu His Gly Thr Thr Ala 20 25 30 Thr Phe His Arg Cys Ala Lys Asp Pro Trp Arg Leu Pro Gly Thr Tyr 35 40 45 Val Val Val Leu Lys Glu Glu Thr His Leu Ser Gln Ser Glu Arg Thr 50 55 60 Ala Arg Arg Leu Gln Ala Gln Ala Ala Arg Arg Gly Tyr Leu Thr Lys 65 70 75 80 Ile Leu His Val Phe His Gly Leu Leu Pro Gly Phe Leu Val Lys Met 85 90 95 Ser Gly Asp Leu Leu Glu Leu Ala Leu Lys Leu Pro His Val Asp Tyr 100 105 110 Ile Glu Glu Asp Ser Ser Val Phe Ala Gln Ser Ile Pro Trp Asn Leu 115 120 125 Glu Arg Ile Thr Pro Pro Arg Tyr Arg Ala Asp Glu Tyr Gln Pro Pro 130 135 140 Asp Gly Gly Ser Leu Val Glu Val Tyr Leu Leu Asp Thr Ser Ile Gln 145 150 155 160 Ser Asp His Arg Glu Ile Glu Gly Arg Val Met Val Thr Asp Phe Glu 165 170 175 Asn Val Pro Glu Glu Asp Gly Thr Arg Phe His Arg Gln Ala Ser Lys 180 185 190 Cys Asp Ser His Gly Thr His Leu Ala Gly Val Val Ser Gly Arg Asp 195 200 205 Ala Gly Val Ala Lys Gly Ala Ser Met Arg Ser Leu Arg Val Leu Asn 210 215 220 Cys Gln Gly Lys Gly Thr Val Ser Gly Thr Leu Ile Gly Leu Glu Phe 225 230 235 240 Ile Arg Lys Ser Gln Leu Val Gln Pro Val Gly Pro Leu Val Val Leu 245 250 255 Leu Pro Leu Ala Gly Gly Tyr Ser Arg Val Leu Asn Ala Ala Cys Gln 260 265 270 Arg Leu Ala Arg Ala Gly Val Val Leu Val Thr Ala Ala Gly Asn Phe 275 280 285 Arg Asp Asp Ala Cys Leu Tyr Ser Pro Ala Ser Ala Pro Glu Val Ile 290 295 300 Thr Val Gly Ala Thr Asn Ala Gln Asp Gln Pro Val Thr Leu Gly Thr 305 310 315 320 Leu Gly Thr Asn Phe Gly Arg Cys Val Asp Leu Phe Ala Pro Gly Glu 325 330 335 Asp Ile Ile Gly Ala Ser Ser Asp Cys Ser Thr Cys Phe Val Ser Gln 340 345 350 Ser Gly Thr Ser Gln Ala Ala Ala His Val Ala Gly Ile Ala Ala Met 355 360 365 Met Leu Ser Ala Glu Pro Glu Leu Thr Leu Ala Glu Leu Arg Gln Arg 370 375 380 Leu Ile His Phe Ser Ala Lys Asp Val Ile Asn Glu Ala Trp Phe Pro 385 390 395 400 Glu Asp Gln Arg Val Leu Thr Pro Asn Leu Val Ala Ala Leu Pro Pro 405 410 415 Ser Thr His Gly Ala Gly Trp Gln Leu Phe Cys Arg Thr Val Trp Ser 420 425 430 Ala His Ser Gly Pro Thr Arg Met Ala Thr Ala Val Ala Arg Cys Ala 435 440 445 Pro Asp Glu Glu Leu Leu Ser Cys Ser Ser Phe Ser Arg Ser Gly Lys 450 455 460 Arg Arg Gly Glu Arg Met Glu Ala Gln Gly Gly Lys Leu Val Cys Arg 465 470 475 480 Ala His Asn Ala Phe Gly Gly Glu Gly Val Tyr Ala Ile Ala Arg Cys 485 490 495 Cys Leu Leu Pro Gln Ala Asn Cys Ser Val His Thr Ala Pro Pro Ala 500 505 510 Glu Ala Ser Met Gly Thr Arg Val His Cys His Gln Gln Gly His Val 515 520 525 Leu Thr Gly Cys Ser Ser His Trp Glu Val Glu Asp Leu Gly Thr His 530 535 540 Lys Pro Pro Val Leu Arg Pro Arg Gly Gln Pro Asn Gln Cys Val Gly 545 550 555 560 His Arg Glu Ala Ser Ile His Ala Ser Cys Cys His Ala Pro Gly Leu 565 570 575 Glu Cys Lys Val Lys Glu His Gly Ile Pro Ala Pro Gln Glu Gln Val 580 585 590 Thr Val Ala Cys Glu Glu Gly Trp Thr Leu Thr Gly Cys Ser Ala Leu 595 600 605 Pro Gly Thr Ser His Val Leu Gly Ala Tyr Ala Val Asp Asn Thr Cys 610 615 620 Val Val Arg Ser Arg Asp Val Ser Thr Thr Gly Ser Thr Ser Glu Gly 625 630 635 640 Ala Val Thr Ala Val Ala Ile Cys Cys Arg Ser Arg His Leu Ala Gln 645 650 655 Ala Ser Gln Glu Leu Gln Ser Gly Ser Gly Gly Leu Asn Asp Ile Phe 660 665 670 Glu Ala Gln Lys Ile Glu Trp His Glu Asn Leu Tyr Phe Gln Gly His 675 680 685 His His His His His 690

Claims (71)

Compounds of formula I:
[Formula I]

(In the formula,
A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR ⁶ , and —C(O)NR ⁶ R ⁷ ;
B is selected from H, alkyl, and halo, or
A and B together with the carbon atom to which they are attached form a 5 or 6 membered heteroaryl;
X is NR ⁵ or O;
R ¹ and R ^1′ are each independently selected from H and alkyl; or
when n is 0, R ¹ and R ^1′ together with the atoms to which they are attached form a 4 to 8 membered cycloalkyl or cycloalkenyl ring;
R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or
R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring; or
R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;
R ^2′ is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano, or
R ² and R ^2′ together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;
each of R ³ and R ⁴ is independently H or alkyl; or
R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring;
R ⁵ is H or alkyl; or
R ¹ and R ⁵ together with the atoms to which they are attached form a 6 to 8 membered cycloalkyl or heterocyclyl ring; or
R ² and R ⁵ together with the atoms to which they are attached form a 5 to 8 membered cycloalkyl or heterocyclyl ring;
each R ⁶ and R ⁷ is independently H or alkyl;
Y is selected from aryl, heteroaryl and heterocyclyl;
n is 0 or 1). 2. The method of claim 1, wherein A is selected from H, hydroxy, thioalkyl, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR ⁶ , and -C(O)NR ⁶ R ⁷ . compound. The compound of claim 1 , wherein A is H. The compound of claim 1 , wherein A is alkyl. The compound of claim 1 , wherein A is thioalkyl. The compound of claim 1 , wherein A is alkoxy. The compound of claim 1 , wherein A is cycloalkyl. The compound of claim 1 , wherein A is selected from -SCH ₃ , -SCHF ₂ , and -OCHF _{2 .} The compound of claim 1 , wherein A and B together with the carbon atom to which they are attached form a pyrrolyl or thienyl ring, which is unsubstituted or substituted with one or more alkyl. 10. The compound according to any one of claims 1 to 9, wherein B is H. 11. A compound according to any one of claims 1 to 10, wherein X is NR ^{5 .} 12. A compound according to any one of claims 1 to 11, wherein Y is heteroaryl or heterocyclyl. 13. A compound according to any one of claims 1 to 12, wherein R ¹ and R ¹ ′ are each H. 13. The method according to any one of claims 1 to 12, wherein n is 0 and R ¹ and R ^{1 ′} together with the atoms to which they are attached are unsubstituted or substituted with 4 to eg one or more hydroxy. which form an 8 membered monocyclic or bicyclic cycloalkyl or cycloalkenyl ring. 15. The compound of claim 14, wherein R ¹ and R ^1' together with the atoms to which they are attached form a 4 to 8 membered monocyclic or bicyclic cycloalkyl. 15. The compound of claim 14, wherein R ¹ and R ^1' together with the atoms to which they are attached form a 4 to 8 membered monocyclic or bicyclic cycloalkenyl. 15. The compound of claim 14, wherein the cycloalkyl ring is a cyclopentyl ring. 15. The compound of claim 14, wherein the cycloalkyl ring is substituted with hydroxyl or hydroxyalkyl. 19. The compound of any one of claims 1-18, wherein R ² is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, alkylamino, cyano, and hydroxyl. 20. The compound of any one of claims 1-19, wherein R ² is C _1-3 alkyl. 21. The compound of claim 20, wherein R ² is substituted with one or more substituents selected from amino, amido, cyano, hydroxy, and heterocyclyl. 22. The compound of any one of claims 1-21, wherein R ^2' is C _1-3 alkyl. 22. The compound of any one of claims 1-21, wherein R ^2' is H. 13. A compound according to any one of claims 1 to 12, wherein R ¹ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. 13. A compound according to any one of claims 1 to 12, wherein R ^1′ and R ² together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. 13. A compound according to any one of claims 1 to 12, wherein R ² and R ^2' together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. 27. The compound of any one of claims 1-26, wherein R ³ is C _1-3 alkyl. 27. The compound of any one of claims 1-26, wherein R ³ is H. 13. A compound according to any one of claims 1 to 12, wherein R ² and R ³ together with the atoms to which they are attached form a 3 to 8 membered cycloalkyl or heterocyclyl ring. 30. The compound of any one of claims 1-29, wherein R ⁴ is H. 13. The compound of any one of claims 1-12, wherein R ¹ and R ⁵ together with the atoms to which they are attached form a 6-8 membered cycloalkyl or heterocyclyl ring. 13. A compound according to any one of claims 1 to 12, wherein R ² and R ⁵ together with the atoms to which they are attached form a 5 to 8 membered cycloalkyl or heterocyclyl ring. 33. The compound of any one of claims 1-32, wherein Y is monocyclic heteroaryl. 34. The compound of claim 33, wherein Y is selected from pyridyl, pyridinyl, pyrazinyl, pyrimidinyl, and thiazolyl. 34. The compound of claim 33, wherein Y is selected from trizenyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl and triazolyl. 36. A compound according to any one of claims 33 to 35, wherein monocyclic heteroaryl is unsubstituted, or alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, aryl, heteroaryl; substituted with one or more substituents selected from heterocyclyl, nitro, sulfonamido, and thioalkyl. 37. The method of claim 36, wherein the monocyclic heteroaryl is phenyl, pyridinyl, 2-hydroxypyridinyl, piperidinonyl, 2-hydroxy-1-methylpyridinyl, triazolyl, imidazolidinonyl, pyrimido substituted with a 6-membered aryl, heteroaryl or heterocyclyl selected from nyl, 2-hydroxyisoquinolinyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinonyl . 38. The method of claim 37, wherein the monocyclic heteroaryl is substituted with heteroaryl or heterocyclyl substituted with one or more substituents selected from halo, CN, alkyl, alkoxy, hydroxy, carboxy, -CO _{2 alkyl, and tetrazolyl} compound that is. 37. The compound of claim 36, wherein the monocyclic heteroaryl is disposed in the para position of A with respect to X. 33. The compound of any one of claims 1-32, wherein Y is bicyclic heteroaryl. 41. The compound of claim 40, wherein Y is selected from benzothiazolyl, benzoxazolyl, benzimidazolyl, triazolopyridinyl, thiazolopyridinyl, quinolinyl, and quinoxalinyl. 42. The method of claim 40 or 41, wherein the bicyclic heteroaryl is unsubstituted, or alkyl, haloalkyl, hydroxyalkyl, thioalkyl, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamido substituted with one or more substituents. 42. The compound of claim 40 or 41, wherein the bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from thioalkyl, alkoxycarbonyl, amido, carboxy, halo, and heteroaryl. 44. The method of any one of claims 1 to 43, wherein Y is substituted with an amido substituent ^{of the formula -C(O)NR 8} R ⁹ or -NR ⁹ C(O)R ^{10 , wherein:}
R ⁸ and R ⁹ are each independently selected from H, alkyl, heterocyclyl and heteroaryl; or
R ⁸ and R ⁹ together with the nitrogen atom to which they are attached form a 4, 5, 6, or 7 membered heterocyclic or heteroaryl ring;
R ¹⁰ is alkyl. 43. The method of any one of claims 1-42, wherein Y is substituted with a sulfonamido substituent _{of the formula -S(O) 2} NR ⁸ R ⁹ or -NR ⁹ S(O) ₂ R ^{10 , wherein}
R ⁸ and R ⁹ are each independently selected from H, alkyl, and heteroaryl; or
R ⁸ and R ⁹ together with the nitrogen atom to which they are attached form a 4, 5, 6, or 7 membered heterocyclic ring;
R ¹⁰ is alkyl. 46. The compound of claim 44 or 45, wherein R ⁸ and R ⁹ are each independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl. 47. The method of any one of claims 44-46, wherein ^{one or both of R 8} and R ⁹ are alkyl and each alkyl is independently unsubstituted or methyl, methoxy, carboxy, cyano, hydroxy , dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-di substituted with one or more substituents selected from hydro-3H-1,2,4-triazol-3-onyl. 46. The method of claim 44 or 45, wherein R ⁸ and R ⁹ together with the nitrogen atom to which they are attached are aziradine, isothiazolidin-1,1-dioxide, azetidine, thiazol-4(5Hn)-one , morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane. 46. The method of claim 44 or 45, wherein R ⁸ and R ⁹ together with the nitrogen atom to which they are attached are 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine , octahydropyrazino [2,1-c] [1,4] oxazine, tetrahydropyrido [3,4-d] pyrimidine, 2-oxa-8-azaspiro [4.5] decane, tetrahydropyr Rolo[3,4-c]pyrazole, thiomorpholine, 2-oxa-7-azaspiro[3.5]nonane, 2,8-diazaspiro[4.5]decan-3-one, tetrahydro-1,7 -naphthyridine, 1-oxa-4,9-diazaspiro [5.5] undecan-3-one, tetrahydropyrrolo [3,4-d] imidazole, pyrimidine, 8-oxa-2-azaspiro [4.5]decane, hexahydro-3H-oxazolo[3,4-a]pyrazin-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro- [1,2,4]triazolo[4,3-a]pyrazine, 2,7-diazaspiro[4.4]nonane, 2,6-diazaspiro[3.4]octane, 7-oxa-2-azaspiro [3.5] nonane, 1-oxa-8λ ² -azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridinone, tetrahydro-[ 1,2,4]triazolo[4,3-a]pyrazin-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7- A compound which forms a heterocyclic ring selected from diazaspiro[3.5]nonane, pyrazole, and pyridazin-3(2H)-one. 49. The compound of claim 48, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl. 49. The compound of claim 48, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxy, and oxetanyl. A compound selected from:

53. A pharmaceutical composition comprising a compound according to any one of claims 1-52 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. 53. A method of treating a cardiovascular disease in a subject comprising administering to the subject a compound according to any one of claims 1-52. 55. The method of claim 54, wherein the cardiovascular disease is selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome and coronary artery disease. how it is. 56. The method of claim 54 or 55, wherein the cardiovascular disease is familial hypercholesterolemia. 56. The method of claim 54 or 55, wherein the cardiovascular disease is autosomal dominant hypercholesterolemia. 58. The method of any one of claims 54-57, wherein the level of circulating serum cholesterol is reduced in the subject. 59. The method of any one of claims 54-58, wherein the level of circulating serum LDL-cholesterol is reduced in the subject. 60. The method of any one of claims 54-59, wherein the level of circulating serum VLDL-cholesterol is reduced in the subject. 61. The method of any one of claims 54 to 60, wherein the level of circulating serum triglycerides is reduced in the subject. 62. The method of any one of claims 54-61, wherein the level of circulating serum lipoprotein A is reduced in the subject. 63. The method of any one of claims 54-62, wherein the subject has atherosclerosis. 64. The method of any one of claims 54-63, wherein atherosclerotic plaque formation is reduced in the subject. 65. The method of any one of claims 54-64, wherein the subject has a gain- of-function mutation in the PCSK9 gene. 66. The method of any one of claims 54-65, further comprising co-administering one or more additional therapeutic agents. 67. The method of claim 66, wherein the one or more additional therapeutic agents is alirocumab, evolocumab, bococizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimbe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluva statin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin. 67. The method of claim 66, wherein the one or more additional therapeutic agents are HMG-CoA reductase inhibitors, HMG-CoA synthetase inhibitors, HMG-CoA reductase gene expression inhibitors, HMG-CoA synthetase gene expression inhibitors, MTP/apo B secretion inhibitors, CETP inhibitor, bile acid absorption inhibitor, cholesterol absorption inhibitor, cholesterol synthesis inhibitor, squalene synthase inhibitor, squalene epoxidase inhibitor, squalene cyclase inhibitor, squalene epoxidase/squalene cyclase combination inhibitor, fibrate, niacin, niacin and lovastatin combination , an ion exchange resin, an antioxidant, an ACAT inhibitor, a bile acid sequestrant and a PCSK9 translation inhibitor. 53. A method of treating sepsis or septic shock in a subject comprising administering to the subject a compound according to any one of claims 1-52. 70. The method of claim 69, further comprising co-administering one or more additional therapeutic agents. 71. The method of claim 70, wherein the one or more additional therapeutic agents is kanamycin, amikacin, tobramycin, dibecacin, gentamicin, sisomycin, netylmycin, streptomycin, neomycin B, C and E, imiphenum, meropenum , Ertapenem, Doripenum, Panifenum, Viapenem, Rasufenum, Tebifenum, Lenafenum, Tomophenum, Tienphenum, Ciprofloxacin, Garenoxacin, Gatifloxacin, Gemifloxacin, Levofloxacin, Sinoxacin, Nalidec Acid, moxifloxacin, oxolinic acid, pyromidic acid, pipemidic acid, rosoxacin, enoxacin, pleloxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, lufloxacin , valofloxacin, grepafloxacin, levofloxacin, pajufloxacin, spafloxacin, temfloxacin, tosufloxacin, clinafloxacin, cytafloxacin, trovafloxacin, prulifloxacin, derafloxacin , JNJ-Q2, nemonoxacin, zabofloxacin, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, metacycline, minocycline, rolytetracycline, tigecycline, and ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol and ticarcillin.